KR101783994B1 - A method for diagnosing a gastric cancer and a diagnostic kit using the method - Google Patents

Abstract

The present invention relates to a method for providing information for diagnosis of gastric cancer and a diagnostic kit for gastric cancer using the same. The present inventors measured the gene mutation of gastric cancer using semiconductor based sequencing of nucleic acid isolated from formalin-fixed paraffin-embedded (FFPE) tissue samples and analyzed copy number variation using nCounter, / It is possible to diagnose gastric cancer cost-effectively. In particular, in the present invention, it is possible to diagnose stomach cancer more accurately through analysis of a gene set optimized for stomach cancer diagnosis.

Description

TECHNICAL FIELD The present invention relates to a gastric cancer diagnosis method and a gastric cancer diagnostic kit using the same,

The present invention relates to a diagnostic method for gastric cancer and a diagnostic kit for gastric cancer using the same, and more particularly, to a method for diagnosing gastric cancer using semiconductor-based sequencing and copy number change (CNV) analysis in a formalin-fixed paraffin-embedded (FFPE) And a diagnostic kit.

Gastric cancer is the fourth most common cancer worldwide and the second leading cause of death [1], with a high incidence in Asian countries including Korea [2]. Recent treatment options for stomach cancer have been identified and available for physicians and patients [3-5].

In such targeted therapies, mutation profiling of the causative cancer is important. Mutant profiling can also be performed using Sanger sequencing, but is costly, time-consuming, and labor-intensive, making it unsuitable for clinical applications. In addition, Sanger sequencing requires a lot of DNA, and it is impossible to analyze several genes at the same time using a small amount of tissue sample. Next generation sequencing (NGS) solves this problem using multiplexing, a high-efficiency sequencing method capable of simultaneously analyzing multiple genes in multiple samples [6, 7]. One of the NGS platforms, the Ion Torrent AmpliSeq Cancer Panel, relies on the non-optical detection of hydrogen ions in semiconductor devices [8], which can detect 2,855 carcinogenic mutations on 50 mutant genes. This method is superior to other mass spectroscopy-based sequencing methods and provides faster sequencing results at lower cost [8]. Formalin-fixed paraffin-embedded (FFPE) tissue samples containing small amounts of DNA can be applied to this sequencing method. The Ion Torrent AmpliSeq Cancer Panel has shown high sensitivity to known carcinogenic mutations [9,10] and is the method used in the five major US cancer centers for molecular diagnosis of target therapy [11] .

Amplification of oncogenes is a major mechanism of gene overexpression and contributes to tumorigenesis [12]. For example, HER2, MET, FGFR2 and KRAS genes are overexpressed in stomach cancer [13,14]. FISH (Fluorescent in situ hybridization) and / or IHC (immunohistochemistry) are widely used for the detection of copy number variations (CNV) of clinical samples. However, high cost and biopsy are limited due to the small sample size, and high sensitivity, low cost, and high-efficiency techniques are required for easier access. The nCounter CNV CodeSets (Nanostring technologies, Life Sciences, Seattle, WA) are excellent in accuracy and reproducibility and provide excellent and fast results with less effort than real-time quantitative PCR or CNV arrays [15].

An excellent customized cancer treatment method will improve the patient's outcome. A sample of the patient's tumor is needed to characterize the cancer at the molecular level and to identify subgroups of the disease that require different treatments. FFPE tissue is important for this study [16]. Recordings for Individual Customized Target Treatments To determine if AmpliSeq and nCounter custom CNV panels could be applied to clinical samples, we analyzed FFPE gastric cancer samples using AmpliSeq and nCounter custom CNV panels.

Numerous papers and patent documents are referenced and cited throughout this specification. The disclosures of the cited papers and patent documents are incorporated herein by reference in their entirety to better understand the state of the art to which the present invention pertains and the content of the present invention.

The present inventors sought to develop a novel stomach cancer diagnostic method using formalin-fixed paraffin-embedded (FFPE) tissue samples. As a result, the inventors measured the mutation of the gastric cancer-associated gene using semiconductor based sequencing of the nucleic acid isolated from the formalin-fixed paraffin-embedded (FFPE) tissue sample and analyzed the copy number variation using nCounter, It is possible to diagnose cancer more time / cost effectively. In addition, "TP53, APC, PIK3CA, KRAS, SMO, STK11, CDKN2A and SMAD4" as a highly accurate gene set for diagnosing gastric cancer; HER2, CCNE1, MYC, KRAS, and EGFR "as a gene set for analysis of CNVs were identified, and it was confirmed that diagnosis of gastric cancer could be performed more accurately through analysis of a gene set optimized for diagnosis of gastric cancer, thereby completing the present invention .

Accordingly, an object of the present invention is to provide a method for providing information for diagnosis of gastric cancer.

Another object of the present invention is to provide a gastric cancer diagnostic kit.

Other objects and advantages of the present invention will become more apparent from the following detailed description of the invention, claims and drawings.

According to one aspect of the present invention, the present invention provides a method for screening CNV (gastric cancer-associated marker gene) of a gastric cancer-associated molecular marker gene using nucleic acids isolated from formalin-fixed paraffin-embedded (FFPE) The present invention provides a method for diagnosing and treating cancer of the stomach cancer, comprising the steps of:

The present inventors sought to develop a novel stomach cancer diagnostic method using formalin-fixed paraffin-embedded (FFPE) tissue samples. As a result, the inventors measured the mutation of the gastric cancer-associated gene using semiconductor based sequencing of the nucleic acid isolated from the formalin-fixed paraffin-embedded (FFPE) tissue sample and analyzed the copy number variation using nCounter, It is possible to diagnose cancer more time / cost effectively. HER2, CCNE1, MYC, KRAS, and EGFR, which are gene sets for analysis of CNVs, and "TP53, APC, PIK3CA, KRAS, SMO, STK11, CDKN2A and SMAD4" And that the diagnosis of gastric cancer can be performed more accurately through the analysis of the gene set optimized for diagnosis of gastric cancer, thereby completing the present invention.

In tailored target therapy, mutant profiling of cancer is important in determining the treatment method. The use of formalin-fixed paraffin-embedded (FFPE) tissue is important to characterize cancer at the molecular level. We performed AmpliSeq Cancer Hotspot Panel v2 and nCounter Copy Number Variation Assay on 89 FFPE stomach cancer samples to determine if this method could be applied to clinical samples for personalized target therapy. The results were verified by Sanger sequencing, real-time quantitative PCR, fluorescence in situ hybridization (FISH) and IHC (immunohistochemistry). The somatic mutations in TP53 (28.17%), APC 10.1%, PIK3CA 5.6%, KRAS 4.5%, SMO 3.4%, STK11 3.4%, CDKN2A 3.4% and SMAD4 3.4% Was observed; Amplification of the HER2, CCNE1, MYC, KRAS and EGFR genes in 8 (8.9%), 4 (4.5%), 2 (2.2%), 1 (1.1%) and 1 A phenomenon was observed. In the case of the gene amplified samples, HER 2 gene amplification was confirmed in FISH and IHC, and EGFR and CCNE1 were also confirmed to be overexpressed in tumor cells. That is, the present invention successfully performed semiconductor-based sequencing and nCounter copy number analysis of FFPE gastric cancer samples.

Sanger sequencing has been used for gene mutation profiling for cancer diagnosis in the past, but time and cost have been greatly reduced due to the development of the next generation sequencing method. On the other hand, FISH and IHC have been widely used in the past to detect copy number variation, but they are expensive and have a disadvantage in that the amount of samples is insufficient. With this in mind, the inventors of the present invention have developed a high-efficiency screening method of clinical samples capable of analyzing hotspot mutation and gene amplification more quickly, accurately, and cost-effectively in relation to cancer-related genes.

The term " subject to be diagnosed " in the present invention includes a subject already suffering from gastric cancer or a subject susceptible to stomach cancer, and may be a mammal. Preferred mammals include, but are not limited to, humans, non-human primates, mice, rats, dogs, cats, horses, and cows.

In order to carry out the method of the present invention, a biological sample can be collected from a subject to be diagnosed, from which a paraffin block is prepared using the gastric cancer tissue and the normal tissue.

The FFPE tissue sample can provide a stable nucleic acid used for CNV analysis, and the nucleic acid sample can be isolated through a conventional method known in the art or a commercially available extraction kit or the like.

The method of the present invention will be described in detail as follows:

First, the CNV (copy number variation) analysis of the gastric cancer-associated molecular marker gene is performed with the nucleic acid isolated from the FFPE tissue sample.

The information providing method of the present invention provides a method for diagnosing gastric cancer by performing CNV analysis of a gastric cancer-associated molecular marker gene.

According to a specific embodiment of the present invention, in the present invention, purified genomic DNA is applied to nCounter Copy Number Variation CodeSets for CNV analysis. That is, in the present invention, the CNV analysis was performed using the "NanoString nCounter analyzing system", and the study using the NanoString was fast, sensitive, and highly reproducible.

According to one embodiment of the present invention, the gene to be analyzed is human epidermal growth factor receptor (HER2), CCNE1 (cyclin E1), viral myelocytoma viral oncogene homolog (MYC), epidermal growth factor receptor (EGFR) (Kirsten rat sarcoma viral oncogene homolog). HER2, CCNE1, MYC, EGFR and KRAS, which are the genes to be analyzed according to the present invention, can be selected from the sequences of SEQ ID NO 1 (Acc No. NM_004448), SEQ ID No. 2 (Acc No. NM_001238), SEQ ID No. 3 (NM_002467), Sequence Listing 4 (Acc No. NM_005228) and Sequence Listing 5 (Acc No. NM_004985). CNV analysis is performed on at least one gene selected from the group consisting of the above-mentioned genes. More specifically, the HER2 is a target sequence selected from among SEQ ID NOS: 6 to 8, and CCNE1 is a target sequence selected from SEQ ID NOS: 9 to 11, And MYC is a target sequence selected from SEQ ID NO: 12 to SEQ ID NO: 14, EGFR is a target sequence selected from SEQ ID NO: 15 to SEQ ID NO: 17, KRAS is a sequence selected from SEQ ID NO: One or more sequences selected from the 18th to 20th sequences are referred to as target sequences.

According to another embodiment of the present invention, the CNVs of the genes are characterized by the following numerical ranges:

The genetic CNV range of SEQ ID NO: 1 sequence is 5-65;

The gene CNV range of SEQ ID NO: 2 sequence is 5-25;

The genetic CNV range of the third sequence is 10-40;

The gene CNV range of the fourth sequence is 1-10; And

The genomic CNV range of SEQ ID NO: 5 sequence is 10-25.

According to a particular embodiment of the present invention, the CNVs of the genes are characterized by the following numerical ranges:

The gene CNV range of SEQ ID NO: 1 sequence 9-62;

The genomic CNV range of the sequence of SEQ ID No. 2 is 8-22;

The genomic CNV range of SEQ ID NO: 3 sequence 13-38;

The gene CNV range of the sequence of SEQ ID No. 4 is 6-9; And

As the CNV range of the gene of Sequence Listing 5 sequence 17-19.

The CNV analysis result can be confirmed by directly measuring the expression rate of the gene. Gene amplification of HER2, CCNE1, MYC, EGFR and KRAS can be confirmed by measuring the expression level of mRNA level or protein level thereof. For example, RT-PCR, northern blotting, , Microarray, NPA (nuclease protection assay (NPA), or in situ hybridization) can be used. However, as a method for measuring the protein level, , Immunohistochemistry, ELISA, radioimmunoassay, radioimmunoprecipitation, Ouchterlony immunodiffusion, rocket immunoelectrophoresis, IHC, FISH, immunoprecipitation assays, complement fixation, FACS, and protein chips , But is not limited thereto.

The expression levels of HER2, CCNE1, MYC, EGFR and KRAS were measured as described above. When the expression levels of HER2, CCNE1, MYC, EGFR and KRAS were increased compared with normal specimens, it was judged that the copy number of the gene was increased.

According to one embodiment of the present invention, the gastric cancer is gastric adenocarcinoma, and it can be used in other types of cancer.

According to one embodiment of the present invention, the information providing method of the present invention is considered to increase the probability of becoming a target treatment target of personalized treatment when the frequency of the gastric cancer-related gene mutation is high and the gastric cancer-associated gene CNVs is high .

Meanwhile, the information providing method of the present invention may further include a step of performing sequencing of the separated nucleic acid to measure a mutation of a gastric cancer-associated molecular marker. In this step, sequencing of nucleic acids isolated from formalin-fixed paraffin-embedded (FFPE) tissue samples of the subject to be diagnosed is performed to detect the mutation of the gastric cancer-associated molecular markers .

The gene to be sequenced may be selected from the group consisting of TP53 (tumor protein 53), adenomatous polyposis coli (APC), phosphatidylinositol 4,5-bisphosphate 3-kinase catalytic subunit alpha, KRAS (Kirsten rat sarcoma viral oncogene homolog), SMO Is one or more genes selected from the group consisting of STK11 (serine-threonine kinase 11), CDKN2A (cyclin-dependent kinase inhibitor 2A) and SMAD4 (SMAD family member 4).

The sequencing method is a next generation sequencing (NGS) method, and more particularly, a semiconductor-based sequencing method that is one of the NGS platforms. According to a particular embodiment of the present invention, the sequencing method is an Ion Torrent AmpliSeq Cancer Panel. As a method that relies on the non-optical detection of protons in semiconductor devices, the present inventors have used this to detect 2,855 carcinogenic mutations on 50 genes. This method is superior to other mass spectrometry-based sequencing methods and can provide faster sequencing results at lower cost. IonTorrent Software can be used to perform automatic data analysis of sequencing results.

The gene that can be analyzed in the above step is gastric cancer-related genes such as TP53 (tumor protein 53), adenomatous polyposis coli (APC), PIK3CA (phosphatidylinositol 4,5-bisphosphate 3-kinase catalytic subunit alpha), KRAS (Kirsten rat sarcoma viral oncogene homolog, SMO (smoothened), serine-threonine kinase 11 (STK11), cyclin-dependent kinase inhibitor 2A (CDKN2A) and SMAD4 (SMAD family member 4).

The genes to be analyzed, TP53, APC, PIK3CA, KRAS, SMO, STK11, CDKN2A and SMAD4 are shown in Sequence Listing 21 (Acc No. NP_000537.3; 393 aa), Sequence Listing 22 (Acc No. NP_000029. 2, 2843 aa), SEQ ID NO: 23 (Acc No. NP_006209.1; 1068 aa), SEQ ID NO: 24 (Acc No. NP_004976.2; 188 aa), SEQ ID NO: 25 (Acc No. NP_000446.1; 433 aa), SEQ ID NO: NP_005622.1 (78 No. aa), Sequence Listing 26 sequence (Acc No. NP_000446.1; 433 aa) 28 sequence (Acc No. NP_005350.1; 552 aa); The amino acid sequences are shown in SEQ ID NO: 29 (Acc No. NM_000546; 2591 bp), SEQ ID NO: 30 (ACC No. NM_000038; 10740 bp), SEQ ID NO: 31 (Acc No. NM_006218.1; 3424 bp (Acc No. NM_004985; 5765 bp), SEQ ID NO: 33 (Acc No. NM_005631.3; 3772 bp), SEQ ID NO: 34 (Acc No. NM_000455: 3286 bp), SEQ ID NO: Is encoded by the nucleotide sequence of List 35 sequence (Acc No. NM_000077; 1267 bp) and SEQ ID No. 36 sequence (Acc No. NM_005359.3; 3220 bp).

On the other hand, the above-mentioned gene mutation can lead to a change of the following amino acid sequence:

(I) TP53: amino acid changes of the sequence of SEQ ID NO: 21, R248Q, R248W, R213fs * 34, R213 *, R273H, R273C, R175H, R185R, R342 *, C135C, C135fs * 35, C176S, D208V, G245R, T626C , L206fs * 41, V173A or Y236C;

(Ii) APC: K1359E, K1363E or P1433L as an amino acid change of the second sequence of the sequence listing;

(Iii) PIK3CA: E545K, N1044K, E1037K or H1047R as amino acid changes of SEQ ID NO: 23;

(Iv) KRAS: G13V or G12V as an amino acid change in SEQ ID NO: 24;

(V) SMO: E518K, E208K or R512H as amino acid changes of Sequence Listing No. 25;

(Vi) STK11: amino acid change of SEQ ID NO: 26 sequence, S31F or T32I;

(Ⅶ) CDKN2A: amino acid change of SEQ ID NO: 27 sequence, T79I, H66R or C315A; or

(Ⅷ) SMAD4: Amino acid change of SEQ ID NO: 28, R361H, M447I or Q448X.

The method may further include performing a manual review step using the cutoff value of the frequency and coverage of the gene mutation after measuring the gene mutation. Through this manual review process, data of higher quality can be obtained. According to a specific embodiment of the present invention, an automatic mutation-invocation algorithm is used to detect a predicted mutation from sequencing results, and a manual review uses a cutoff value of at least 6% variance and x100 coverage.

According to another aspect of the present invention, there is provided a diagnostic kit for stomach cancer using nucleic acid isolated from formalin-fixed paraffin-embedded (FFPE) tissue samples of a subject to be diagnosed, The present invention provides a stomach cancer diagnostic kit characterized in that it is judged to be a target treatment target of an immunotherapeutic treatment in which a copy number variation (CNV) of a molecular marker gene appears.

Since the kit of the present invention uses an information providing method for diagnosis of the gastric cancer target described above, the description common to both of them is omitted in order to avoid the excessive complexity of the present specification.

The features and advantages of the present invention are summarized as follows:

(a) The present invention relates to a method for providing information for diagnosis of gastric cancer and a diagnostic kit for gastric cancer using the same.

(b) The present inventors analyzed nucleic acid isolated from formalin-fixed paraffin-embedded (FFPE) tissue samples and analyzed copy number variation using nCounter, which proved that it is possible to diagnose gastric cancer in a more cost effective and time- .

(c) Especially in the present invention, it is possible to diagnose stomach cancer more accurately through analysis of gene set optimized for stomach cancer diagnosis.

FIG. 1 schematically shows a variant call processing.
Figure 2 shows immunohistochemical staining of EGFR, CCNE1 and HER2, and FISH results of HER2. Expression of EGFR (A), CCNE1 (B), and HER2 (C) was very strong in samples with increased copy number. The IHC results showed that the samples with HER2 2+ showed amplification of the HER2 gene in FISH (D).
Figure 3 shows the coverage of the Ion AmpliSeq v2 cancer panel and Oncomap v4.
Figure 4 is a graphical representation of the relationship between the MET CNVs detected by nCounter and the mRNA level of the MET gene exhibited by real-time PCR.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are only for describing the present invention in more detail and that the scope of the present invention is not limited by these embodiments in accordance with the gist of the present invention .

Example

Experimental material

Sample

In the microscopic examination, the tumor cells were excised from the 4 mm non-stained fragment in an amount of 75% or more compared with the H & E staining slides. The Qiagen DNA FFPE Tissue Kit (Qiagen, Hilden, Germany) DNA was extracted. After DNA extraction, the concentration and the ratio at 260/280 nm and 260/230 nm were measured using a spectrophotometer (ND1000, Nanodrop Technologies, Thermo-Fisher Scientific, MA, USA). Each sample was quantitated with a Qubit fluorescence system (Life Technologies, Carlsbad, Calif.). Ten nanograms of genomic DNA were measured by Qubit fluorescence spectrometry and used for library production. Seven samples were excluded from the experiment due to failure of library production. Finally, a sample of 89 (31 female, 58 male) samples were analyzed. Table 1 shows the clinical and pathological characteristics of the patients. During the interim evaluation period of 76 months, recurrence or metastasis occurred in 11 patients (range 5.5-149.3). This study was approved by the IRB of the Samsung Medical Center and conducted in accordance with the principles set forth in the Helsinki Declaration. IRB did not receive informed consent because of retrospective analysis and anonymity of data. Samples were collected by general methods and all information was anonymous.

Clinical pathological features of 89 gastric cancer patients n = 89 gender female 31 male 58 age Average 53 Middle age 55 Lauren Classification Intestinal type 27 Distributed type 60 Mixed type 2 part Top 1/3 11 Middle part 1/3 32 Lower 1/3 46 pT step T1,2 23 T3 51 T4 15 pN step N1 41 N2 44 N3 4 AJCC / UICC stage (7 ^th ed) Ⅰ One Ⅱ 22 Ⅲ 66 Relapse and / or distant metastasis existence 11 Nonexistence 78 Subsequent cycle (month) Medium value (range) 76 (5.5-149.3)

Ion AmpliSeq cancer panel v2

Using the Ion AmpliSeq Cancer Panel v2 (Ion Torrent), the present inventors analyzed selected somatic mutations based on a literature review. As a result, a total of 2855 mutations were detected in 50 mutagenic and tumor suppressor genes. First, 10 ng of DNA was extracted from each of 89 FFPE tumor samples and multiplex PCR amplification was performed in a single tube using Ion Ampli-SeqCancer Primer Pool and Ion AmpliSeqKit reagent (Life Technologies). The amplification product, the amplification product, was treated with FuPa reagent to partially cleave the primer and phosphorylate the amplicon. After ligation of the phosphorylated ampicillin with Ion Adapters, it was purified. For barcoded library production, the non-barcoded adapter mix was replaced with a bar codeed adapter of the Ion Xpress Barcode Adapters 1-96 kit. The ligated DNA underwent nicktranslation and amplification procedures to complete the connection between the adapter and the amplicon, thereby creating a material for downstream molding. The input DNA and unreacted primers were removed through two Agencourt AMPure XP reagent binding reactions at 0.6 and 1.2 bead / sample volume ratios. The final library molecule was 125,300 bp in size. Next, the present inventors applied the library to the Ion OneTouch System for the production of an automated mold. Sequencing was performed using Ion PGM sequencer and automatic data analysis was performed using IonTorrent Software.

To determine the sensitivity and specificity of the Ion AmpliSeq cancer panel, we used the entire exome sequencing results of four stomach cancer samples with mutation data identified [17].

nCounter Copy Number Variation CodeSets

To analyze the copy number change (CNV), 300 ng of purified genomic DNA was extracted from 2-3 fragments of a 4 mm thick FFPE tumor block using QIAamp DNA FFPE tissue kit (Qiagen, Hilden, Germany) And then applied to the nCounter Copy Number Variation CodeSets. The DNA was denatured at 95 ° C after treatment with AluI restriction enzyme. Using the nCounter Cancer CN assay kit (Nanostring Technologies), the segmented DNA and the code set of 86 genes were hybridized at 65 ° C for 18 hours. The signals of the reporter probes were measured with nCounter analyzer and tabulated. The average count number was greater than or equal to 3 and the results were reconfirmed by IHC, FISH or real time PCR.

HER2 , EGFR  ( HER1 ) And CCNE1 Immunohistochemical study of IHC )

To verify the CNV results from nCounter, IHC of HER2 was performed on all samples and IHC of EGFR and CCNE1 was performed on some samples. After removing paraffin and water, 4 mm sections were immunostained with HER 2 on silane-coated slides. HercepTest (Dako, Glostrup, Denmark) was performed as described in the prior art [18]. (1: 100 dilution; Novocastra / Vision Biosystems, Newcastle, UK) was used for EGFR staining and anti-CCNE1 / Cyclin E1 antibody HE12; 1: 200 dilution; Thermo Fisher Scientific, MA). Ventana BenchMark XT was used for automatic slide processing. The results were analyzed by an expert pathologist.

HER2 of Fluorescent nucleic acid hybridization  Experiment( Fluorescence in situ hybridization )

FISH was performed using a two-color DNA-specific probe of PathVision ™ (Abbott / Vysis: LSI HER2 SpectrumOrange ™ and CEP 17 SpectrumGreen ™) [19]. We counted the hybridization signal in 20 nuclei per sample under a fluorescence microscope (Zeiss Axioskop). Nested nuclei were excluded and measured only when present as a single nucleus. The HER2 gene is considered to be amplified when the FISH signal ratio of HER2 / CEP17 is 2.0 or more [20].

KRAS  And MET  Real time for amplification PCR

First, DNA was obtained from FFPE gastric cancer tumor tissue. The PCR reaction mixture consisted of 2 μL genomic DNA template, 10 μL Taqman Universal PCR Master Mix (Applied Biosystems Inc, Foster City, CA) and 0.2 μM primers, respectively. To detect the correct CN change, one site in intron 1 (TaqMan Copy Number Assay Hs06943812_cn), one site in intron 2 (Hs002534878_cn), three different sites on the KRAS gene; And one region (Hs02739788_cn) in exon 6 were analyzed. The MET gene used a primer previously used by the present inventors [21]: 1 part in exon 3-intron 3 (TaqMan Copy Number Assay Hs01602615_cn), 1 part in intron 12 (Hs0527935_cn); And one site (Hs02884964_cn) in intron 20-exon 21

On the other hand, the copy number gain was measured using the following profiling: 50 캜 2 min; 95 ° C 10 min denaturation; 40 cycles - 95 캜 for 15 seconds, 60 캜 for 1 minute. The relative quantification of the results of experiment 4 was then determined using the 7900 HT fast real-time PCR system. RNaseP assay kit (Applied Biosystems) was used in the control group. After the amplification reaction, the results of the experiment with the critical-cycle value for the copy number and the result of the reference analysis were obtained for analysis of post-PCR data and analyzed by CopyCaller software (Applied Biosystems) [22]. Based on the results of at least two of the three probes, the CN gain state and the KRAS copy number were assigned.

Analysis method

An automated mutation-invocation algorithm was used to detect predicted mutations, and the same changes that occurred thereafter were excluded. Recurrent calls were observed in more than 10 of the 89 samples and were classified as false positive. In order to detect the mutation change [9,10] according to the inventors' previous studies, a cutoff value of more than 6% mutation rate and x100 coverage was used. After reviewing each polymorphism in the list of somatic mutations in cancer (COSMIC, http://cancer.sanger.ac.uk/cancergenome/projects/cosmic), a single-nucleotide polymorphism was extracted (Fig. 1). For the mutant genes (TP53, APC, PIK3CA, STK11, CDKN2A, KRAS, HRAS, BRAF and CTNNB1) known in gastric cancer, the results of automatic invocation were reviewed to obtain deletion mutants with somewhat lower variability.

Experiment result

Ion AmpliSeq cancer panel Result of

A total of 8189 mutations were observed from 89 samples, of which 3554 were classified as non-identity changes. After filtering out recurrent calls, a variation of less than 6% - an allele incidence, coverage of less than 100X and the list at the intron site, 65 of these variant calls were selected. Additionally, automated calls were examined in known mutations such as BRAF, KRAS and PIK3CA, and two mutation signals were stored and excluded during the filtering process. Of the 89 samples, 39 (43.8%) had at least one mutation (Figure 1). Twenty-two and five mutations were found in two samples, respectively. The MLH1 somatic mutation [missense mutation in exon 20: c.1147A.G (p.M383 V)] was observed in the 5 mutant samples, and MLH1 protein was not expressed by the MLH1 promoter and methylation. IHC [23] Respectively. This result means that the sample is a hypermutated tumor sample. On the other hand, samples with 22 mutations were not able to reconfirm the same results through Sanger sequencing, despite having passed the rate and coverage cutoff values. This is a false positive result due to DNA purity. Therefore, samples with 22 mutations were excluded from the final analysis. (5 cases, 5.6%), KRAS (3 cases, 3.4%), SMO (4 cases, 4.5%), STK11 (24 cases, 27.0% 3 cases, 3.4%), somatic cell mutations including CDKN2A (3 cases, 3.4%) and SMAD4 (3 cases, 3.4%) were detected (Table 2 and Table 3). Table 2 and Table 3 also show amino acid changes in mutant genes. We observed in 21 patients (21.3%) that two or more specific somatic mutations or multiple somatic mutations occurred.

On the other hand, somatic mutations of TP53, ERBB4 and CTNNB1 without false-positive signals were identified using four samples of stomach cancer with known mutation rates through whole milk full sequencing (data not shown).

Mutation and amino acid change frequency in 89 gastric cancer samples -1 gene N * % Amino acid change (N,%) TP53 24 27.0 R248Q (N = 3, 3.3%) ^§ R248W (N = 1, 1.1%) R213fs * 34 (N = 1, 1.1%) R213 * (N = 1, 1.1%) R273H (N = 1, 1.1%) R273C (N = 1, 1.1%) R175H (N = 2, 2.2%) R185R (N = 1, 1.1%) R342 * (N = 1, 1.1%) C135C (N = 1, 1.1%) C135fs * 35 (N = 2, 2.2%) C176S (N = 1, 1.1%) D208V (N = 1, 1.1%) G245R (N = 1, 1.1%) T626C (N = 1, 1.1%) L206fs * 41 (N = 1, 1.1%) V173A (N = 1, 1.1%) Y236C (N = 1, 1.1%) APC 9 10.1 K1359E (N = 1, 1.1%) ^§ K1363E (N = 1, 1.1%) P1433L (N = 1, 1.1%) PIK3CA 5 5.6 E545K (N = 2, 2.2%) N1044K (N = 1, 1.1%) E1037K (N = 1, 1.1%) H1047R (N = 1, 1.1%) KRAS 3 3.4 G13V (N = 2, 2.2%) G12V (N = 1, 1.1%) SMO 3 3.4 E518K (N = 1, 1.1%) E208K (N = 1, 1.1%) R512H (N = 1, 1.1%) STK11 3 3.4 S31F (N = 1, 1.1%) ^§ T32I (N = 1, 1.1%)

Mutation and amino acid change frequency in 89 gastric cancer samples -2 gene N * % Amino acid change (N,%) CDKN2A 3 3.4 T79I (N = 1, 1.1%) H66R (N = 1, 1.1%) C315A (N = 1, 1.1%) SMAD4 3 3.4 R361H (N = 1, 1.1%) M447I (N = 1, 1.1%) Q448X (N = 1, 1.1%) CDH1 2 2.2 L343P (N = 1, 1.1%) G382D (N = 1, 1.1%) FBXW7 2 2.2 R393 * (N = 1, 1.1%) ^§ ATM One 1.1 Y861H (N = 1, 1.1%) CTNNB1 One 1.1 T41A (N = 1, 1.1%) ERBB2 One 1.1 V842I (N = 1, 1.1%) FGFR2 One 1.1 G906A (N = 1, 1.1%) FGFR3 One 1.1 A369A (N = 1, 1.1%) KDR One 1.1 W1143X (N = 1, 1.1%) MLH1 One 1.1 A424G (N = 1, 1.1%) PTEN One 1.1 R15S (N = 1, 1.1%) SMARCB1 One 1.1 H177Y (N = 1, 1.1%) RET One 1.1 A641T (N = 1, 1.1%)

* N: Number of samples with mutations, §INS / DEL in the remaining cases.

nCounter  The amplification reaction and IHC , FISH  Or real time PCR Verification by

(8.9%), four (4.5%), two (2.2%), one (1.1%) and one (1.1%) samples of HER2, CCNE1, MYC, KRAS and EGFR gene amplification (Table 4). The amplification phenomenon of the MET, FGFR2, CDK4 and CDK6 genes was not observed in any samples. In the case of the sample in which the amplification reaction was observed, protein overexpression was also observed in the immunohistochemical staining experiments of HER2, EGFR and CCNE1 (Fig. 2A, B and C). FISH experiments on samples with HER2 2+ in HercepTest revealed heterologous amplification of the HER2 gene (Figure 2D).

In a real-time PCR experiment of KRAS, we observed an increase in the copy number in one sample (36, 37 and 49); In the samples without KRAS amplification, the copy number did not increase (0.9 - 2.4, mean 1.4).

Change of the copy number in gastric cancer samples observed with nCounter gene Number of samples with CNV CNV range (average) Of the 89 samples, ERBB2 * 8 (8.9%) 9-62 (31.9) CCNE1 4 (4.5%) 8-22 (12.8) MYC 2 (2.2%) 13-38 (25.5) EGFR 1 (1.1%) 7 KRAS 1 (1.1%) 18

* All samples were positive in immunohistochemical staining (8 samples: 3+; 1 sample: 2+; 1 sample: 1+).

Because of its rarity [21], no positive samples were found for the MET gene. Thus, we used 10 stomach cancer samples (5 amplified samples and 5 unamplified samples) and MET amplified gastric cancer cell lines (MKN45 and SNU5) that were known to have a copy number and mRNA amount. CNVs detected by nCounter were highly correlated with the copy number (data not shown) and MET gene mRNA levels detected by real-time PCR (r = 0.874, p = 0.001) (Fig.

Argument

Using Ion AmpliSeq v2, we found somatic mutations in 39 out of 89 samples, which means that these platforms can easily be applied to FFPE tissue samples. TP53 was the most frequent mutation, followed by APC, PIK3CA and KRAS. In addition, CN increase in HER2, CCNE1, MYC, EGFR and KRAS genes was successfully detected using the CNV panel used in the present invention, and these results were verified by IHC and real-time PCR.

The mutation frequencies in the COSMIC database were found to be very similar to the data obtained in the present invention: TP53 (32%), PIK3CA (10%), KRAS (6%), APC (6%), CTNNB1 5%), FBXW7 (5%), SMO (4%), ERBB2 (2%) and STK11 (2%). Recent total sequencing studies of gastric adenocarcinoma have shown a higher incidence of mutations in TP53 (36% and 73%) and PIK3CA (14% and 20%) compared to our results [24,25] . Compared with the results of whole-genuin sequencing, the frequency of mutation was low in this study, but the present inventors' mass spectrometer-based OncoMap v4. [26] Compared to data, this is a significant increase. Both AmpliSeq and OncoMap are able to detect mutations at critical sites and explain why mutation frequency is low in oncogenes and tumor suppressor genes. Fig. 3 shows the results of comparing detection mutation profiles of AmpliSeq v2 and OncoMap v4.

Onco-map experiments with 237 gastric cancer samples showed a higher PIK3CA mutation in the late stage patients (5.1% in Stage IV; 6.4% in stage II / III; 2.4% in stage IB) [26]. In the present invention, three PIK3CA mutations occurred in stage Ⅲ patients and two mutations occurred in stage Ⅱ. These results support their biological role in tumor progression. On the other hand, mutation of HER2 (ERBB2) c.2524G> A (V842I) was observed in one sample. In preclinical studies, cell lines with the V842I mutation were resistant to trastuzumab and sensitive to neratinib, an irreversible HER2 inhibitor [27].

Compared to mass spectrometer-based sequencing, semiconductor-based sequencing has differences in detection and signal conversion. Instead of using optical methods to detect nucleotide changes, they detect changes in pH. In other words, nucleotides are inserted during DNA strand formation, which detects pH changes due to proton (H +) release [8]. Therefore, the cost and time for data processing is greatly reduced compared to other NGS platforms. It is very important that patients with invasive cancer, including gastric cancer patients, can obtain information about mutations quickly and accurately at low cost.

In the present invention, after applying cutoff values of frequency and coverage, manually reviewing the automatic signals in known mutations and then adding two signals that exhibit low side coverage. Although these coverage values did not reach initial baseline values, the measured frequency values were higher than the initial setting values (6%) and the quality of the data was also excellent. This is why the importance of manual review after the automatic screening is emphasized. A recently published study has used AmpliSeq for platform analysis, which also emphasizes the benefits of manual review-based screening data [9,10].

Personalized targeted therapies for terminal cancer patients mainly rely on "oncogene (or oncogene) addiction." In other words, the maintenance of survival and survival of tumor cells can be accomplished by using one or more genes, [29] The open-label, international, clinical Phase III, randomized controlled trial of ToGa (Trastuzumab for Gastric Cancer) showed that the combination of trastuzumab and chemotherapy was a new standard for HER2-positive end stage or gastric cancer patients [29] Preclinical studies have demonstrated the efficacy of EGFR or MET amplification and overexpression of gastric cancers in the treatment of cetuximab or MET receptor tyrosine kinase inhibitors [30]. Dependent kinase inhibition [31]. In the case of targeted therapy with high-efficiency technology, FISH and array analysis methods using conventional genomic hybridization are low-cost, high-cost, and laborious and time-consuming methods in the genomic region [32]. The nCounter CNV assay method , The present invention has been shown to be applicable to FFPE clinical samples and validated as a result of IHC, FISH or real-time PCR. Although not verified for all genes, The results of genetic testing are noteworthy.

In summary, we have successfully performed semiconductor-based sequencing and nCounter CNV assays in 89 gastric cancer FFPE tissue samples. High-throughput sequencing and CNV screening can be used to detect hotspot mutations and CNV in genes more quickly, accurately, and cost-effectively. In a personalized genomic medicine era, it is expected that targeted therapies for gastric cancer patients will be possible using these techniques.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the same is by way of illustration and example only and is not to be construed as limiting the scope of the present invention. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

references

1. Jemala, Bray F, Center MM, Ferlay J, Ward E, et al. (2011) Global cancer statistics. CA Cancer J Clin 61: 6990.

2. Shin A, Kim J, Park S (2011) Gastric cancer epidemiology in Korea. J Gastric Cancer 11: 135140.

3. Smyth EC, Cunningham D (2012) Targeted therapy for gastric cancer. Curr Treat Options Oncol 13: 377389.

4. Wong H, Yau T (2013) Molecular targeted therapies in advanced gastric cancer: does tumor histology matter? Therap Adv Gastroenterol 6: 1531.

5. Nadauld LD, Ford JM (2013) Molecular profiling of gastric cancer: toward personalized cancer medicine. J Clin Oncol 31: 838839.

6. Metzker ML (2010) Sequencing technologies - the next generation. Nat Rev Genet 11: 3146.

7. MacConaill LE, Campbell CD, Kehoe SM, Bass AJ, Hatton C, et al. (2009) Profiling critical cancer gene mutations in clinical tumor samples. PLoS One 4: e7887.

8. Rothberg JM, Hinz W, Rearick TM, Schultz J, Mileski W, et al. (2011) An integrated semiconductor device enabling non-optical genome sequencing.

Nature 475: 348352.

9. Singh RR, Patel KP, Routbort MJ, Reddy NG, Barkoh BA, et al. (2013) Clinical validation of a next-generation sequencing screen for mutational hotspots in 46 cancer-related genes. J Mol Diagn 15: 607622.

10. Beadling C, Neff TL, Heinrich MC, Rhodes K, Thornton M, et al. (2013) Combining highly multiplexed PCR with semiconductor-based sequencing for rapid cancer genotyping. J Mol Diagn 15: 171176.

11. Simon R, Roychowdhury S (2013) Implementing personalized cancer genomics in clinical trials. Nat Rev Drug Discov 12: 358369.

12. Dai Z, Zhu WG, Morrison CD, Brena RM, Smiraglia DJ, et al. (2003) A comprehensive search for DNA amplification in lung cancer identifies inhibitors of apoptosis cIAP1 and cIAP2 as candidate oncogenes. Hum MoI Genet 12: 791801.

13. Wu Y, Grabsch H, Ivanova T, Tan IB, Murray J, et al. (2013) Comprehensive genomic meta-analysis identifies intra-tumoral stroma as a predictor of survival in patients with gastric cancer. Gut 62: 11001111.

14. Deng N, Goh LK, Wang H, Das K, Tao J, et al. (2012) A comprehensive survey of genomic alterations in gastric cancer reveals systematic patterns of molecular exclusivity and co-occurrence among distinct therapeutic targets. Gut 61: 673684.

15. Geiss GK, Bumgarner RE, Birditt B, Dahl T, Dowidar N, et al. (2008) Direct multiplexed measurement of gene expression with color-coded probe pairs. Nat Biotechnol 26: 317325.

16. Austin Tanney RDK (2010) Developing mRNA-based biomarkers from formalin-fixed paraffin-embedded tissue. Future Medicine 7: 205211.

17. Kang G, Hwang WC, Do IG, Wang K, Kang SY, et al. (2013) Exome sequencing identifies early gastric carcinoma as an early stage of advanced gastric cancer. PLoS One 8: e82770.

18. Cho EY, Park K, Do I, Cho J, Kim J, et al. (2013) Heterogeneity of ERBB2 in gastric carcinomas: a study of tissue microarray and matched primary and metastatic carcinomas. Mod Pathol 26: 677684.

19. Cho EY, Srivastava A, Park K, Kim J, Lee MH, et al. (2012) Comparison of four immunohistochemical tests and FISH for measuring HER2 expression in gastric carcinomas. Pathology 44: 216220.

20. Ruschoff J, Hanna W, Bilous M, Hofmann M, Osamura RY, et al. (2012) HER2 testing in gastric cancer: a practical approach. Mod Pathol 25: 637650.

21. Ha SY, Lee J, Kang SY, Do IG, Ahn S, et al. (2013) MET overexpression assessed by new interpretation method predicts gene amplification and poor survival in advanced gastric carcinomas. Mod Pathol 26: 16321641.

22. Graziano F, Galluccio N, Lorenzini P, Ruzzo A, Canestrari E, et al. (2011) Genetic Activation of the MET Pathway and Prognosis of Patients with High-Risk, Radically Resected Gastric Cancer. J Clin Oncol.

23. Lee SE, Kang SY, Cho J, Lee B, Chang DK, et al. (2014) Pyloric gland adenoma in Lynch syndrome. Am J Surg Pathol 38: 784792.

24. Wang K, Kan J, Yuen ST, Shi ST, Chu KM, et al. (2011) Exome sequencing identifies frequent mutations of ARID1A in molecular subtypes of gastric cancer. Nat Genet 43: 12191223.

25. Zang ZJ, Cutcutache I, Poon SL, Zhang SL, McPherson JR, et al. (2012) Exome sequencing of gastric adenocarcinoma identifies recurrent somatic mutations in cell adhesion and chromatin remodeling genes. Nat Genet 44: 570574.

26. Lee J, van Hummelen P, Go C, Palescandolo E, Jang J, et al. (2012) Highthroughput mutation profiling identifies frequent somatic mutations in advanced gastric adenocarcinoma. PLoS One 7: e38892.

27. Bose R, Kavuri SM, Searleman AC, Shen W, Shen D, et al. (2013) Activating HER2 mutations in HER2 gene amplification negative breast cancer. Cancer Discov 3: 224237.

28. Ma PC (2012) Personalized targeted therapy in advanced non-small cell lung cancer. Cleve Clin J Med 79 Electronic Suppl 1: eS5660.

29. Bang YJ, Van Cutsem E, Feyereislovaya, Chung HC, Shen L, et al. HER2-positive advanced gastric or gastro-oesophageal junction cancer (TOGA): a phase 3, open-label, randomized controlled trial. Lancet 376: 687697.

30. Zhang L, Yang J, Cai J, Song X, Deng J, et al. (2013) A subset of gastric cancers with EGFR amplification and overexpression respond to cetuximab therapy. Sci Rep 3: 2992.

31. Network TCGAR (2014) Comprehensive molecular characterization of gastric adenocarcinoma. Nature doi: 10.1038 / nature13480.

32. Duan J, Zhang JG, Deng HW, Wang YP (2013) Comparative studies of copy number variation detection methods for next-generation sequencing technologies. PLoS One 8: e59128.

<110> SAMSUNG LIFE PUBLIC WELFARE FOUNDATION <120> A method for diagnosing a gastric cancer and a diagnostic kit          using the method <130> PN150001 <160> 36 <170> Kopatentin 2.0 <210> 1 <211> 4664 <212> DNA <213> homo sapiens_HER2 gene <400> 1 gcttgctccc aatcacagga gaaggaggag gtggaggagg agggctgctt gaggaagtat 60 aagaatgaag ttgtgaagct gagattcccc tccattggga ccggagaaac caggggagcc 120 ccccgggcag ccgcgcgccc cttcccacgg ggccctttac tgcgccgcgc gcccggcccc 180 cacccctcgc agcaccccgc gccccgcgcc ctcccagccg ggtccagccg gagccatggg 240 gccggagccg cagtgagcac catggagctg gcggccttgt gccgctgggg gctcctcctc 300 gccctcttgc cccccggagc cgcgagcacc caagtgtgca ccggcacaga catgaagctg 360 cggctccctg ccagtcccga gacccacctg gacatgctcc gccacctcta ccagggctgc 420 caggtggtgc agggaaacct ggaactcacc tacctgccca ccaatgccag cctgtccttc 480 ctgcaggata tccaggaggt gcagggctac gtgctcatcg ctcacaacca agtgaggcag 540 gtcccactgc agaggctgcg gattgtgcga ggcacccagc tctttgagga caactatgcc 600 ctggccgtgc tagacaatgg agacccgctg aacaatacca cccctgtcac aggggcctcc 660 ccaggaggcc tgcgggagct gcagcttcga agcctcacag agatcttgaa aggaggggtc 720 ttgatccagc ggaaccccca gctctgctac caggacacga ttttgtggaa ggacatcttc 780 cacaagaaca accagctggc tctcacactg atagacacca accgctctcg ggcctgccac 840 ccctgttctc cgatgtgtaa gggctcccgc tgctggggag agagttctga ggattgtcag 900 agcctgacgc gcactgtctg tgccggtggc tgtgcccgct gcaaggggcc actgcccact 960 gactgctgcc atgagcagtg tgctgccggc tgcacgggcc ccaagcactc tgactgcctg 1020 gcctgcctcc acttcaacca cagtggcatc tgtgagctgc actgcccagc cctggtcacc 1080 tacaacacag acacgtttga gtccatgccc aatcccgagg gccggtatac attcggcgcc 1140 agctgtgtga ctgcctgtcc ctacaactac ctttctacgg acgtgggatc ctgcaccctc 1200 gtctgccccc tgcacaacca agaggtgaca gcagaggatg gaacacagcg gtgtgagaag 1260 tgcagcaagc cctgtgcccg agtgtgctat ggtctgggca tggagcactt gcgagaggtg 1320 agggcagtta ccagtgccaa tatccaggag tttgctggct gcaagaagat ctttgggagc 1380 ctggcatttc tgccggagag ctttgatggg gacccagcct ccaacactgc cccgctccag 1440 ccagagcagc tccaagtgtt tgagactctg gaagagatca caggttacct atacatctca 1500 gcatggccgg acagcctgcc tgacctcagc gtcttccaga acctgcaagt aatccgggga 1560 cgaattctgc acaatggcgc ctactcgctg accctgcaag ggctgggcat cagctggctg 1620 gggctgcgct cactgaggga actgggcagt ggactggccc tcatccacca taacacccac 1680 ctctgcttcg tgcacacggt gccctgggac cagctctttc ggaacccgca ccaagctctg 1740 ctccacactg ccaaccggcc agaggacgag tgtgtgggcg agggcctggc ctgccaccag 1800 ctgtgcgccc gagggcactg ctggggtcca gggcccaccc agtgtgtcaa ctgcagccag 1860 ttccttcggg gccaggagtg cgtggaggaa tgccgagtac tgcaggggct ccccagggag 1920 tatgtgaatg ccaggcactg tttgccgtgc caccctgagt gtcagcccca gaatggctca 1980 gtgacctgtt ttggaccgga ggctgaccag tgtgtggcct gtgcccacta taaggaccct 2040 cccttctgcg tggcccgctg ccccagcggt gtgaaacctg acctctccta catgcccatc 2100 tggaagtttc cagatgagga gggcgcatgc cagccttgcc ccatcaactg cacccactcc 2160 tgtgtggacc tggatgacaa gggctgcccc gccgagcaga gagccagccc tctgacgtcc 2220 atcatctctg cggtggttgg cattctgctg gtcgtggtct tgggggtggt ctttgggatc 2280 ctcatcaagc gacggcagca gaagatccgg aagtacacga tgcggagact gctgcaggaa 2340 acggagctgg tggagccgct gacacctagc ggagcgatgc ccaaccaggc gcagatgcgg 2400 atcctgaaag agacggagct gaggaaggtg aaggtgcttg gatctggcgc ttttggcaca 2460 gtctacaagg gcatctggat ccctgatggg gagaatgtga aaattccagt ggccatcaaa 2520 gtgttgaggg aaaacacatc ccccaaagcc aacaaagaaa tcttagacga agcatacgtg 2580 atggctggtg tgggctcccc atatgtctcc cgccttctgg gcatctgcct gacatccacg 2640 gtgcagctgg tgacacagct tatgccctat ggctgcctct tagaccatgt ccgggaaaac 2700 cgcggacgcc tgggctccca ggacctgctg aactggtgta tgcagattgc caaggggatg 2760 agctacctgg aggatgtgcg gctcgtacac agggacttgg ccgctcggaa cgtgctggtc 2820 aagagtccca accatgtcaa aattacagac ttcgggctgg ctcggctgct ggacattgac 2880 gagacagagt accatgcaga tgggggcaag gtgcccatca agtggatggc gctggagtcc 2940 attctccgcc ggcggttcac ccaccagagt gatgtgtgga gttatggtgt gactgtgtgg 3000 gagctgatga cttttgggc caaaccttac gatgggatcc cagcccggga gatccctgac 3060 ctgctggaaa agggggagcg gctgccccag ccccccatct gcaccattga tgtctacatg 3120 atcatggtca aatgttggat gattgactct gaatgtcggc caagattccg ggagttggtg 3180 tctgaattct cccgcatggc cagggacccc cagcgctttg tggtcatcca gaatgaggac 3240 ttgggcccag ccagtccctt ggacagcacc ttctaccgct cactgctgga ggacgatgac 3300 atgggggacc tggtggatgc tgaggagtat ctggtacccc agcagggctt cttctgtcca 3360 gaccctgccc cgggcgctgg gggcatggtc caccacaggc accgcagctc atctaccagg 3420 agtggcggtg gggacctgac actagggctg gagccctctg aagaggaggc ccccaggtct 3480 ccactggcac cctccgaagg ggctggctcc gatgtatttg atggtgacct gggaatgggg 3540 gcagccaagg ggctgcaaag cctccccaca catgacccca gccctctaca gcggtacagt 3600 gaggacccca cagtacccct gccctctgag actgatggct acgttgcccc cctgacctgc 3660 agcccccagc ctgaatatgt gaaccagcca gatgttcggc cccagccccc ttcgccccga 3720 gagggccctc tgcctgctgc ccgacctgct ggtgccactc tggaaaggcc caagactctc 3780 tccccaggga agaatggggt cgtcaaagac gtttttgcct ttgggggtgc cgtggagaac 3840 cccgagtact tgacacccca gggaggagct gcccctcagc cccaccctcc tcctgccttc 3900 agcccagcct tcgacaacct ctattactgg gaccaggacc caccagagcg gggggctcca 3960 cccagcacct tcaaagggac acctacggca gagaacccag agtacctggg tctggacgtg 4020 ccagtgtgaa ccagaaggcc aagtccgcag aagccctgat gtgtcctcag ggagcaggga 4080 aggcctgact tctgctggca tcaagaggtg ggagggccct ccgaccactt ccaggggaac 4140 ctgccatgcc aggaacctgt cctaaggaac cttccttcct gcttgagttc ccagatggct 4200 ggaaggggtc cagcctcgtt ggaagaggaa cagcactggg gagtctttgt ggattctgag 4260 gccctgccca atgagactct agggtccagt ggatgccaca gcccagcttg gccctttcct 4320 tccagatcct gggtactgaa agccttaggg aagctggcct gagaggggaa gcggccctaa 4380 gggagtgtct aagaacaaaa gcgacccatt cagagactgt ccctgaaacc tagtactgcc 4440 ccccatgagg aaggaacagc aatggtgtca gtatccaggc tttgtacaga gtgcttttct 4500 gtttagtttt tacttttttt gttttgtttt tttaaagatg aaataaagac ccagggggag 4560 aatgggtgtt gtatggggag gcaagtgtgg ggggtccttc tccacaccca ctttgtccat 4620 ttgcaaatat attttggaaa acagctaaaa aaaaaaaaaa aaaa 4664 <210> 2 <211> 2021 <212> DNA <213> homo sapiens_CCNE1 gene <400> 2 agcagccggc gcggccgcca gcgcggtgta gggggcaggc gcggatcccg ccaccgccgc 60 gcgctcggcc cgccgactcc cggcgccgcc gccgccactg ccgtcgccgc cgccgcctgc 120 cgggactgga gcgcgccgtc cgccgcggac aagaccctgg cctcaggccg gagcagcccc 180 atcatgccga gggagcgcag ggagcgggat gcgaaggagc gggacaccat gaaggaggac 240 ggcggcgcgg agttctcggc tcgctccagg aagaggaagg caaacgtgac cgtttttttg 300 caggatccag atgaagaaat ggccaaaatc gacaggacgg cgagggacca gtgtgggagc 360 cagccttggg acaataatgc agtctgtgca gacccctgct ccctgatccc cacacctgac 420 aaagaagatg atgaccgggt ttacccaaac tcaacgtgca agcctcggat tattgcacca 480 tccagaggct ccccgctgcc tgtactgagc tgggcaaata gagaggaagt ctggaaaatc 540 atgttaaaca aggaaaagac atacttaagg gatcagcact ttcttgagca acaccctctt 600 ctgcagccaa aaatgcgagc aattcttctg gattggttaa tggaggtgtg tgaagtctat 660 aaacttcaca gggagacctt ttacttggca caagatttct ttgaccggta tatggcgaca 720 caagaaaatg ttgtaaaaac tcttttacag cttattggga tttcatcttt atttattgca 780 gccaaacttg aggaaatcta tcctccaaag ttgcaccagt ttgcgtatgt gacagatgga 840 gcttgttcag gagatgaaat tctcaccatg gaattaatga ttatgaaggc ccttaagtgg 900 cgtttaagtc ccctgactat tgtgtcctgg ctgaatgtat acatgcaggt tgcatatcta 960 aatgacttac atgaagtgct actgccgcag tatccccagc aaatctttat acagattgca 1020 gagctgttgg atctctgtgt cctggatgtt gactgccttg aatttcctta tggtatactt 1080 gctgcttcgg ccttgtatca tttctcgtca tctgaattga tgcaaaaggt ttcagggtat 1140 cagtggtgcg acatagagaa ctgtgtcaag tggatggttc catttgccat ggttataagg 1200 gagacgggga gctcaaaact gaagcacttc aggggcgtcg ctgatgaaga tgcacacaac 1260 atacagaccc acagagacag cttggatttg ctggacaaag cccgagcaaa gaaagccatg 1320 ttgtctgaac aaaatagggc ttctcctctc cccagtgggc tcctcacccc gccacagagc 1380 ggtaagaagc agagcagcgg gccggaaatg gcgtgaccac cccatccttc tccaccaaag 1440 acagttgcgc gcctgctcca cgttctcttc tgtctgttgc agcggaggcg tgcgtttgct 1500 tttacagata tctgaatgga agagtgtttc ttccacaaca gaagtatttc tgtggatggc 1560 atcaaacagg gcaaagtgtt ttttattgaa tgcttatagg ttttttttaa ataagtgggt 1620 caagtacacc agccacctcc agacaccagt gcgtgctccc gatgctgcta tggaaggtgc 1680 tacttgacct aagggactcc cacaacaaca aaagcttgaa gctgtggagg gccacggtgg 1740 cgtggctctc ctcgcaggtg ttctgggctc cgttgtacca agtggagcag gtggttgcgg 1800 gcaagcgttg tgcagagccc atagccagct gggcaggggg ctgccctctc cacattatca 1860 gttgacagtg tacaatgcct ttgatgaact gttttgtaag tgctgctata tctatccatt 1920 ttttaataaa gataatactg tttttgaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 1980 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa a 2021 <210> 3 <211> 2379 <212> DNA <213> homo sapiens_MYC gene <400> 3 gacccccgag ctgtgctgct cgcggccgcc accgccgggc cccggccgtc cctggctccc 60 ctcctgcctc gagaagggca gggcttctca gaggcttggc gggaaaaaga acggagggag 120 ggatcgcgct gagtataaaa gccggttttc ggggctttat ctaactcgct gtagtaattc 180 cagcgagagg cagagggagc gagcgggcgg ccggctaggg tggaagagcc gggcgagcag 240 agctgcgctg cgggcgtcct gggaagggag atccggagcg aatagggggc ttcgcctctg 300 gcccagccct cccgctgatc ccccagccag cggtccgcaa cccttgccgc atccacgaaa 360 ctttgcccat agcagcgggc gggcactttg cactggaact tacaacaccc gagcaaggac 420 gcgactctcc cgacgcgggg aggctattct gcccatttgg ggacacttcc ccgccgctgc 480 caggacccgc ttctctgaaa ggctctcctt gcagctgctt agacgctgga tttttttcgg 540 gtagtggaaa accagcagcc tcccgcgacg atgcccctca acgttagctt caccaacagg 600 aactatgacc tcgactacga ctcggtgcag ccgtatttct actgcgacga ggaggagaac 660 ttctaccagc agcagcagca gagcgagctg cagcccccgg cgcccagcga ggatatctgg 720 cgggctctgc 780 tcgccctcct acgttgcggt cacacccttc tcccttcggg gagacaacga cggcggtggc 840 gggagcttct ccacggccga ccagctggag atggtgaccg agctgctggg aggagacatg 900 gtgaaccaga gtttcatctg cgacccggac gacgagacct tcatcaaaaa catcatcatc 960 caggactgta tgtggagcgg cttctcggcc gccgccaagc tcgtctcaga gaagctggcc 1020 tcctaccagg ctgcgcgcaa agacagcggc agcccgaacc ccgcccgcgg ccacagcgtc 1080 tgctccacct ccagcttgta cctgcaggat ctgagcgccg ccgcctcaga gtgcatcgac 1140 ccctcggtgg tcttccccta ccctctcaac gacagcagct cgcccaagtc ctgcgcctcg 1200 caagactcca gcgccttctc tccgtcctcg gattctctgc tctcctcgac ggagtcctcc 1260 ccgcagggca gccccgagcc cctggtgctc catgaggaga caccgcccac caccagcagc 1320 gactctgagg aggaacaaga agatgaggaa gaaatcgatg ttgtttctgt ggaaaagagg 1380 caggctcctg gcaaaaggtc agagtctgga tcaccttctg ctggaggcca cagcaaacct 1440 cctcacagcc cactggtcct caagaggtgc cacgtctcca cacatcagca caactacgca 1500 gcgcctccct ccactcggaa ggactatcct gctgccaaga gggtcaagtt ggacagtgtc 1560 agagtcctga gacagatcag caacaaccga aaatgcacca gccccaggtc ctcggacacc 1620 gaggagaatg tcaagaggcg aacacacaac gtcttggagc gccagaggag gaacgagcta 1680 aaacggagct tttttgccct gcgtgaccag atcccggagt tggaaaacaa tgaaaaggcc 1740 cccaaggtag ttatccttaa aaaagccaca gcatacatcc tgtccgtcca agcagaggag 1800 caaaagctca tttctgaaga ggacttgttg cggaaacgac gagaacagtt gaaacacaaa 1860 cttgaacagc tacggaactc ttgtgcgtaa ggaaaagtaa ggaaaacgat tccttctaac 1920 agaaatgtcc tgagcaatca cctatgaact tgtttcaaat gcatgatcaa atgcaacctc 1980 acaaccttgg ctgagtcttg agactgaaag atttagccat aatgtaaact gcctcaaatt 2040 ggactttggg cataaaagaa cttttttatg cttaccatct tttttttttc tttaacagat 2100 ttgtatttaa gaattgtttt taaaaaattt taagatttac acaatgtttc tctgtaaata 2160 ttgccattaa atgtaaataa ctttaataaa acgtttatag cagttacaca gaatttcaat 2220 cctagtatat agtacctagt attataggta ctataaaccc taattttttt tatttaagta 2280 cattttgctt tttaaagttg atttttttct attgttttta gaaaaaataa aataactggc 2340 aaatatatca ttgagccaaa tcttaaaaaa aaaaaaaaa 2379 <210> 4 <211> 5616 <212> DNA <213> homo sapiens_EGFR gene <400> 4 ccccggcgca gcgcggccgc agcagcctcc gccccccgca cggtgtgagc gcccgacgcg 60 gccgaggcgg ccggagtccc gagctagccc cggcggccgc cgccgcccag accggacgac 120 aggccacctc gtcggcgtcc gcccgagtcc ccgcctcgcc gccaacgcca caaccaccgc 180 gcacggcccc ctgactccgt ccagtattga tcgggagagc cggagcgagc tcttcgggga 240 gcagcgatgc gaccctccgg gacggccggg gcagcgctcc tggcgctgct ggctgcgctc 300 tgcccggcga gtcgggctct ggaggaaaag aaagtttgcc aaggcacgag taacaagctc 360 acgcagttgg gcacttttga agatcatttt ctcagcctcc agaggatgtt caataactgt 420 gaggtggtcc ttgggaattt ggaaattacc tatgtgcaga ggaattatga tctttccttc 480 ttaaagacca tccaggaggt ggctggttat gtcctcattg ccctcaacac agtggagcga 540 attcctttgg aaaacctgca gatcatcaga ggaaatatgt actacgaaaa ttcctatgcc 600 ttagcagtct tatctaacta tgatgcaaat aaaaccggac tgaaggagct gcccatgaga 660 aatttacagg aaatcctgca tggcgccgtg cggttcagca acaaccctgc cctgtgcaac 720 gtggagagca tccagtggcg ggacatagtc agcagtgact ttctcagcaa catgtcgatg 780 gacttccaga accacctggg cagctgccaa aagtgtgatc caagctgtcc caatgggagc 840 tgctggggtg caggagagga gaactgccag aaactgacca aaatcatctg tgcccagcag 900 tgctccgggc gctgccgtgg caagtccccc agtgactgct gccacaacca gtgtgctgca 960 ggctgcacag gcccccggga gagcgactgc ctggtctgcc gcaaattccg agacgaagcc 1020 acgtgcaagg acacctgccc cccactcatg ctctacaacc ccaccacgta ccagatggat 1080 gtgaaccccg agggcaaata cagctttggt gccacctgcg tgaagaagtg tccccgtaat 1140 tatgtggtga cagatcacgg ctcgtgcgtc cgagcctgtg gggccgacag ctatgagatg 1200 gggaagacg gcgtccgcaa gtgtaagaag tgcgaagggc cttgccgcaa agtgtgtaac 1260 ggaataggta ttggtgaatt taaagactca ctctccataa atgctacgaa tattaaacac 1320 ttcaaaaact gcacctccat cagtggcgat ctccacatcc tgccggtggc atttaggggt 1380 gactccttca cacatactcc tcctctggat ccacaggaac tggatattct gaaaaccgta 1440 aaggaaatca cagggttttt gctgattcag gcttggcctg aaaacaggac ggacctccat 1500 gcctttgaga acctagaaat catacgcggc aggaccaagc aacatggtca gttttctctt 1560 gcagtcgtca gcctgaacat aacatccttg ggattacgct ccctcaagga gataagtgat 1620 ggagatgtga taatttcagg aaacaaaaat ttgtgctatg caaatacaat aaactggaaa 1680 aaactgtttg ggacctccgg tcagaaaacc aaaattataa gcaacagagg tgaaaacagc 1740 tgcaaggcca caggccaggt ctgccatgcc ttgtgctccc ccgagggctg ctggggcccg 1800 gagcccaggg actgcgtctc ttgccggaat gtcagccgag gcagggaatg cgtggacaag 1860 tgcaaccttc tggagggtga gccaagggag tttgtggaga actctgagtg catacagtgc 1920 cacccagagt gcctgcctca ggccatgaac atcacctgca caggacgggg accagacaac 1980 tgtatccagt gtgcccacta cattgacggc ccccactgcg tcaagacctg cccggcagga 2040 gtcatgggag aaaacaacac cctggtctgg aagtacgcag acgccggcca tgtgtgccac 2100 ctgtgccatc caaactgcac ctacggatgc actgggccag gtcttgaagg ctgtccaacg 2160 aatgggccta agatcccgtc catcgccact gggatggtgg gggccctcct cttgctgctg 2220 gtggtggccc tggggatcgg cctcttcatg cgaaggcgcc acatcgttcg gaagcgcacg 2280 ctgcggaggc tgctgcagga gagggagctt gtggagcctc ttacacccag tggagaagct 2340 cccaaccaag ctctcttgag gatcttgaag gaaactgaat tcaaaaagat caaagtgctg 2400 ggctccggtg cgttcggcac ggtgtataag ggactctgga tcccagaagg tgagaaagtt 2460 aaaattcccg tcgctatcaa ggaattaaga gaagcaacat ctccgaaagc caacaaggaa 2520 atcctcgatg aagcctacgt gatggccagc gtggacaacc cccacgtgtg ccgcctgctg 2580 ggcatctgcc tcacctccac cgtgcagctc atcacgcagc tcatgccctt cggctgcctc 2640 ctggactatg tccgggaaca caaagacaat attggctccc agtacctgct caactggtgt 2700 gtgcagatcg caaagggcat gaactacttg gaggaccgtc gcttggtgca ccgcgacctg 2760 gcagccagga acgtactggt gaaaacaccg cagcatgtca agatcacaga ttttgggctg 2820 gccaaactgc tgggtgcgga agagaaagaa taccatgcag aaggaggcaa agtgcctatc 2880 aagtggatgg cattggaatc aattttacac agaatctata cccaccagag tgatgtctgg 2940 agctacgggg tgaccgtttg ggagttgatg acctttggat ccaagccata tgacggaatc 3000 cctgccagcg agatctcctc catcctggag aaaggagaac gcctccctca gccacccata 3060 tgtaccatcg atgtctacat gatcatggtc aagtgctgga tgatagacgc agatagtcgc 3120 ccaaagttcc gtgagttgat catcgaattc tccaaaatgg cccgagaccc ccagcgctac 3180 cttgtcattc agggggatga aagaatgcat ttgccaagtc ctacagactc caacttctac 3240 cgtgccctga tggatgaaga agacatggac gacgtggtgg atgccgacga gtacctcatc 3300 ccacagcagg gcttcttcag cagcccctcc acgtcacgga ctcccctcct gagctctctg 3360 agtgcaacca gcaacaattc caccgtggct tgcattgata gaaatgggct gcaaagctgt 3420 cccatcaagg aagacagctt cttgcagcga tacagctcag accccacagg cgccttgact 3480 gaggacagca tagacgacac cttcctccca gtgcctgaat acataaacca gtccgttccc 3540 aaaaggcccg ctggctctgt gcagaatcct gtctatcaca atcagcctct gaaccccgcg 3600 cccagcagag acccacacta ccaggacccc cacagcactg cagtgggcaa ccccgagtat 3660 ctcaacactg tccagcccac ctgtgtcaac agcacattcg acagccctgc ccactgggcc 3720 cagaaaggca gccaccaaat tagcctggac aaccctgact accagcagga cttctttccc 3780 aaggaagcca agccaaatgg catctttaag ggctccacag ctgaaaatgc agaataccta 3840 agggtcgcgc cacaaagcag tgaatttatt ggagcatgac cacggaggat agtatgagcc 3900 ctaaaaatcc agactctttc gatacccagg accaagccac agcaggtcct ccatcccaac 3960 agccatgccc gcattagctc ttagacccac agactggttt tgcaacgttt acaccgacta 4020 gccaggaagt acttccacct cgggcacatt ttgggaagtt gcattccttt gtcttcaaac 4080 tgtgaagcat ttacagaaac gcatccagca agaatattgt ccctttgagc agaaatttat 4140 ctttcaaaga ggtatatttg aaaaaaaaaa aaagtatatg tgaggatttt tattgattgg 4200 ggatcttgga gtttttcatt gtcgctattg atttttactt caatgggctc ttccaacaag 4260 gaagaagctt gctggtagca cttgctaccc tgagttcatc caggcccaac tgtgagcaag 4320 gagcacaagc cacaagtctt ccagaggatg cttgattcca gtggttctgc ttcaaggctt 4380 ccactgcaaa acactaaaga tccaagaagg ccttcatggc cccagcaggc cggatcggta 4440 ctgtatcaag tcatggcagg tacagtagga taagccactc tgtcccttcc tgggcaaaga 4500 agaaacggag gggatggaat tcttccttag acttactttt gtaaaaatgt ccccacggta 4560 cttactcccc actgatggac cagtggtttc cagtcatgag cgttagactg acttgtttgt 4620 cttccattcc attgttttga aactcagtat gctgcccctg tcttgctgtc atgaaatcag 4680 caagagagga tgacacatca aataataact cggattccag cccacattgg attcatcagc 4740 atttggacca atagcccaca gctgagaatg tggaatacct aaggatagca ccgcttttgt 4800 tctcgcaaaa acgtatctcc taatttgagg ctcagatgaa atgcatcagg tcctttgggg 4860 catagatcag aagactacaa aaatgaagct gctctgaaat ctcctttagc catcacccca 4920 accccccaaa attagtttgt gttacttatg gaagatagtt ttctcctttt acttcacttc 4980 aaaagctttt tactcaaaga gtatatgttc cctccaggtc agctgccccc aaaccccctc 5040 cttacgcttt gtcacacaaa aagtgtctct gccttgagtc atctattcaa gcacttacag 5100 ctctggccac aacagggcat tttacaggtg cgaatgacag tagcattatg agtagtgtgg 5160 aattcaggta gtaaatatga aactagggtt tgaaattgat aatgctttca caacatttgc 5220 agatgtttta gaaggaaaaa agttccttcc taaaataatt tctctacaat tggaagattg 5280 gaagattcag ctagttagga gcccaccttt tttcctaatc tgtgtgtgcc ctgtaacctg 5340 actggttaac agcagtcctt tgtaaacagt gttttaaact ctcctagtca atatccaccc 5400 catccaattt atcaaggaag aaatggttca gaaaatattt tcagcctaca gttatgttca 5460 gtcacacaca catacaaaat gttccttttg cttttaaagt aatttttgac tcccagatca 5520 gtcagagccc ctacagcatt gttaagaaag tatttgattt ttgtctcaat gaaaataaaa 5580 ctatattcat ttccactcta aaaaaaaaaa aaaaaa 5616 <210> 5 <211> 5765 <212> DNA <213> homo sapiens_KRAS gene <400> 5 tcctaggcgg cggccgcggc ggcggaggca gcagcggcgg cggcagtggc ggcggcgaag 60 gtggcggcgg ctcggccagt actcccggcc cccgccattt cggactggga gcgagcgcgg 120 cgcaggcact gaaggcggcg gcggggccag aggctcagcg gctcccaggt gcgggagaga 180 ggcctgctga aaatgactga atataaactt gtggtagttg gagctggtgg cgtaggcaag 240 agtgccttga cgatacagct aattcagaat cattttgtgg acgaatatga tccaacaata 300 gaggattcct acaggaagca agtagtaatt gatggagaaa cctgtctctt ggatattctc 360 gacacagcag gtcaagagga gtacagtgca atgagggacc agtacatgag gactggggag 420 ggctttcttt gtgtatttgc cataaataat actaaatcat ttgaagatat tcaccattat 480 agagaacaaa ttaaaagagt taaggactct gaagatgtac ctatggtcct agtaggaaat 540 aaatgtgatt tgccttctag aacagtagac acaaaacagg ctcaggactt agcaagaagt 600 tatggaattc cttttattga aacatcagca aagacaagac agggtgttga tgatgccttc 660 tatacattag ttcgagaaat tcgaaaacat aaagaaaaga tgagcaaaga tggtaaaaag 720 aagaaaaaga agtcaaagac aaagtgtgta attatgtaaa tacaatttgt acttttttct 780 taaggcatac tagtacaagt ggtaattttt gtacattaca ctaaattatt agcatttgtt 840 ttagcattac ctaatttttt tcctgctcca tgcagactgt tagcttttac cttaaatgct 900 tattttaaaa tgacagtgga agtttttttt tcctctaagt gccagtattc ccagagtttt 960 ggtttttgaa ctagcaatgc ctgtgaaaaa gaaactgaat acctaagatt tctgtcttgg 1020 ggtttttggt gcatgcagtt gattacttct tatttttctt accaattgtg aatgttggtg 1080 tgaaacaaat taatgaagct tttgaatcat ccctattctg tgttttatct agtcacataa 1140 atggattaat tactaatttc agttgagacc ttctaattgg tttttactga aacattgagg 1200 gaacacaaat ttatgggctt cctgatgatg attcttctag gcatcatgtc ctatagtttg 1260 tcatccctga tgaatgtaaa gttacactgt tcacaaaggt tttgtctcct ttccactgct 1320 attagtcatg gtcactctcc ccaaaatatt atattttttc tataaaaaga aaaaaatgga 1380 aaaaaattac aaggcaatgg aaactattat aaggccattt ccttttcaca ttagataaat 1440 tactataaag actcctaata gcttttcctg ttaaggcaga cccagtatga aatggggatt 1500 attatagcaa ccattttggg gctatattta catgctacta aatttttata ataattgaaa 1560 agattttaac aagtataaaa aattctcata ggaattaaat gtagtctccc tgtgtcagac 1620 tgctctttca tagtataact ttaaatcttt tcttcaactt gagtctttga agatagtttt 1680 aattctgctt gtgacattaa aagattattt gggccagtta tagcttatta ggtgttgaag 1740 agaccaaggt tgcaaggcca ggccctgtgt gaacctttga gctttcatag agagtttcac 1800 agcatggact gtgtccccac ggtcatccag tgttgtcatg cattggttag tcaaaatggg 1860 gagggactag ggcagtttgg atagctcaac aagatacaat ctcactctgt ggtggtcctg 1920 ctgacaaatc aagagcattg cttttgtttc ttaagaaaac aaactctttt ttaaaaatta 1980 cttttaaata ttaactcaaa agttgagatt ttggggtggt ggtgtgccaa gacattaatt 2040 ttttttttaa acaatgaagt gaaaaagttt tacaatctct aggtttggct agttctctta 2100 acactggtta aattaacatt gcataaacac ttttcaagtc tgatccatat ttaataatgc 2160 tttaaaataa aaataaaaac aatccttttg ataaatttaa aatgttactt attttaaaat 2220 aaatgaagtg agatggcatg gtgaggtgaa agtatcactg gactaggaag aaggtgactt 2280 aggttctaga taggtgtctt ttaggactct gattttgagg acatcactta ctatccattt 2340 cttcatgtta aaagaagtca tctcaaactc ttagtttttt ttttttacaa ctatgtaatt 2400 tatattccat ttacataagg atacacttat ttgtcaagct cagcacaatc tgtaaatttt 2460 taacctatgt tacaccatct tcagtgccag tcttgggcaa aattgtgcaa gaggtgaagt 2520 ttatatttga atatccattc tcgttttagg actcttcttc catattagtg tcatcttgcc 2580 tccctacctt ccacatgccc catgacttga tgcagtttta atacttgtaa ttcccctaac 2640 cataagattt actgctgctg tggatatctc catgaagttt tcccactgag tcacatcaga 2700 aatgccctac atcttatttc ctcagggctc aagagaatct gacagatacc ataaagggat 2760 ttgacctaat cactaatttt caggtggtgg ctgatgcttt gaacatctct ttgctgccca 2820 atccattagc gacagtagga tttttcaaac ctggtatgaa tagacagaac cctatccagt 2880 ggaaggagaa tttaataaag atagtgctga aagaattcct taggtaatct ataactagga 2940 ctactcctgg taacagtaat acattccatt gttttagtaa ccagaaatct tcatgcaatg 3000 aaaaatactt taattcatga agcttacttt ttttttttgg tgtcagagtc tcgctcttgt 3060 cacccaggct ggaatgcagt ggcgccatct cagctcactg caacctccat ctcccaggtt 3120 caagcgattc tcgtgcctcg gcctcctgag tagctgggat tacaggcgtg tgccactaca 3180 ctcaactaat ttttgtattt ttaggagaga cggggtttca ccctgttggc caggctggtc 3240 tcgaactcct gacctcaagt gattcaccca ccttggcctc ataaacctgt tttgcagaac 3300 tcatttattc agcaaatatt tattgagtgc ctaccagatg ccagtcaccg cacaaggcac 3360 tgggtatatg gtatccccaa acaagagaca taatcccggt ccttaggtag tgctagtgtg 3420 gtctgtaata tcttactaag gcctttggta tacgacccag agataacacg atgcgtattt 3480 tagttttgca aagaaggggt ttggtctctg tgccagctct ataattgttt tgctacgatt 3540 ccactgaaac tcttcgatca agctacttta tgtaaatcac ttcattgttt taaaggaata 3600 aacttgatta tattgttttt ttatttggca taactgtgat ttttttagga caattactgt 3660 acacattaag gtgtatgtca gatattcata ttgacccaaa tgtgtaatat tccagttttc 3720 tctgcataag taattaaaat atacttaaaa attaatagtt ttatctgggt acaaataaac 3780 aggtgcctga actagttcac agacaaggaa acttctatgt aaaaatcact atgatttctg 3840 aattgctatg tgaaactaca gatctttgga acactgttta ggtagggtgt taagacttac 3900 acagtacctc gtttctacac agagaaagaa atggccatac ttcaggaact gcagtgctta 3960 tgaggggata tttaggcctc ttgaattttt gatgtagatg ggcatttttt taaggtagtg 4020 gttaattacc tttatgtgaa ctttgaatgg tttaacaaaa gatttgtttt tgtagagatt 4080 ttaaaggggg agaattctag aaataaatgt tacctaatta ttacagcctt aaagacaaaa 4140 atccttgttg aagttttttt aaaaaaagct aaattacata gacttaggca ttaacatgtt 4200 tgtggaagaa tatagcagac gtatattgta tcatttgagt gaatgttccc aagtaggcat 4260 tctaggctct atttaactga gtcacactgc ataggaattt agaacctaac ttttataggt 4320 tatcaaaact gttgtcacca ttgcacaatt ttgtcctaat atatacatag aaactttgtg 4380 gggcatgtta agttacagtt tgcacaagtt catctcattt gtattccatt gatttttttt 4440 ttcttctaaa cattttttct tcaaacagta tataactttt tttaggggat ttttttttag 4500 acagcaaaaa ctatctgaag atttccattt gtcaaaaagt aatgatttct tgataattgt 4560 gtagtaatgt tttttagaac ccagcagtta ccttaaagct gaatttatat ttagtaactt 4620 ctgtgttaat actggatagc atgaattctg cattgagaaa ctgaatagct gtcataaaat 4680 gaaactttct ttctaaagaa agatactcac atgagttctt gaagaatagt cataactaga 4740 ttaagatctg tgttttagtt taatagtttg aagtgcctgt ttgggataat gataggtaat 4800 ttagatgaat ttaggggaaa aaaaagttat ctgcagatat gttgagggcc catctctccc 4860 cccacacccc cacagagcta actgggttac agtgttttat ccgaaagttt ccaattccac 4920 tgtcttgtgt tttcatgttg aaaatacttt tgcatttttc ctttgagtgc caatttctta 4980 ctagtactat ttcttaatgt aacatgttta cctggaatgt attttaacta tttttgtata 5040 gtgtaaactg aaacatgcac attttgtaca ttgtgctttc ttttgtggga catatgcagt 5100 gtgatccagt tgttttccat catttggttg cgctgaccta ggaatgttgg tcatatcaaa 5160 cattaaaaat gaccactctt ttaattgaaa ttaactttta aatgtttata ggagtatgtg 5220 ctgtgaagtg atctaaaatt tgtaatattt ttgtcatgaa ctgtactact cctaattatt 5280 gtaatgtaat aaaaatagtt acagtgacta tgagtgtgta tttattcatg aaatttgaac 5340 tgtttgcccc gaaatggata tggaatactt tataagccat agacactata gtataccagt 5400 gt; tgtaaaggcg tgtttgctta aacttaaaac catatttaga agtagatgca aaacaaatct 5520 gcctttatga caaaaaaata ggataacatt atttatttat ttccttttat caaagaaggt 5580 aattgataca caacaggtga cttggtttta ggcccaaagg tagcagcagc aacattaata 5640 atggaaataa ttgaatagtt agttatgtat gttaatgcca gtcaccagca ggctatttca 5700 aggtcagaag taatgactcc atacatatta tttatttcta taactacatt taaatcatta 5760 ccagg 5765 <210> 6 <211> 100 <212> DNA <213> HER2_CNV target sequence <400> 6 ggccctgagg ttccctcttg acttcagagt cctctccctt cctgtccagc caatgcctgt 60 cttccttttg ggccctacca gcatgacagg gggctgcggg 100 <210> 7 <211> 100 <212> DNA <213> HER2_CNV target sequence <400> 7 gggctcgatg ggcaaagggt ttgagtgaag gcattcatgg tggggagtgg ctggcatggc 60 cagtgctggg agtgatgtcc accctgttcc tggccctgct 100 <210> 8 <211> 100 <212> DNA <213> HER2_CNV target sequence <400> 8 gaattacggg gccacctcag catgtgaagg gagggaaggg gctgcctgtg ccccaccttg 60 cagggtctgt gcacttccca ggattaggga aagaccgggt 100 <210> 9 <211> 100 <212> DNA <213> CCNE1_CNV target sequence <400> 9 agcctgacct cagcaagagt atcagtggaa cattcagtgt aatgataggt ggtgtctcaa 60 tacatttttt tttgactgct tttcttttct ctccttaagg 100 <210> 10 <211> 100 <212> DNA <213> CCNE1_CNV target sequence <400> 10 tcctgttcgc ttcatgaaag cactgagcat ttccagcatt tttgtgtatt aggacctctg 60 tgtggtttat tccctcgaca tgttctccat ctctgaagct 100 <210> 11 <211> 100 <212> DNA <213> CCNE1_CNV target sequence <400> 11 gcccacgatg tcttcactgc agggtatcag tggtgcgaca tagagaactg tgtcaagtgg 60 atggttccat ttgccatggt tataagggag acggggagct 100 <210> 12 <211> 100 <212> DNA <213> MYC_CNV target sequence <400> 12 gttgggtagg cgcaggcagg ggaaaaggga ggcgaggatg tgtccgattc tcctggaatc 60 gttgacttgg aaaaaccagg gcgaatctcc gcacccagcc 100 <210> 13 <211> 100 <212> DNA <213> MYC_CNV target sequence <400> 13 gcggggtggc agggagtgta tgaatgagga taagagagga ttgatctctg agagtgaatg 60 aattgcttcc ctcttaactt ccgagaagtg gtgggattta 100 <210> 14 <211> 100 <212> DNA <213> MYC_CNV target sequence <400> 14 tttgggtgtg tccaaagcct cattaagtct taggtaagaa ttggcatcaa tgtcctatcc 60 tgggaagttg cacttttctt gtccatgcca taacccagct 100 <210> 15 <211> 100 <212> DNA &Lt; 213 > EGFR_CNV target sequence <400> 15 ctcccttcct tgtctgttcc ctcccattgt caggcttctg tggagccata ttcagagcaa 60 catagggagg gggaagagaa aatcaacccc ttggtgaagg 100 <210> 16 <211> 100 <212> DNA &Lt; 213 > EGFR_CNV target sequence <400> 16 gctgcaggtc ccgcaagaca cagagcaacc ctgcaagcca ggtcaccttc cctcttctct 60 gctgtccgac tggccctcca ccatgtgaca ttcaaaagct 100 <210> 17 <211> 100 <212> DNA &Lt; 213 > EGFR_CNV target sequence <400> 17 gcccttccta agactaaatc cagagcactc tgtggtttca gagaagattc ctaaattcca 60 gagtttggac ccagacccag gaattgtgac ttggttggcc 100 <210> 18 <211> 100 <212> DNA <213> KRAS_CNV target sequence <400> 18 ctgtatataa aagattttaa gtgtcaggtg tggtgactaa tgcctgtagt accagcactt 60 tgagagggca agagatgcca ggagtttgag acctcatctc 100 <210> 19 <211> 100 <212> DNA <213> KRAS_CNV target sequence <400> 19 aaaccattct aaagagtaat gaaaaagcag gaggtgggag caaagcacag agtcttagag 60 tcaacccatt acccaggcat ggctacagag catgtaaagc 100 <210> 20 <211> 100 <212> DNA <213> KRAS_CNV target sequence <400> 20 ttggatctta acaccattga ggtttggggt cagataaccc tgttgtgggg gctttcccgt 60 gcactgcagg aagtttaata ttaggttggt gcaaaggtaa 100 <210> 21 <211> 393 <212> PRT <213> Homo sapiens_ TP53 amino acid <400> 21 Met Glu Glu Pro Gln Ser Asp Pro Ser Val Glu Pro Pro Leu Ser Gln   1 5 10 15 Glu Thr Phe Ser Asp Leu Trp Lys Leu Leu Pro Glu Asn Asn Val Leu              20 25 30 Ser Pro Leu Pro Ser Gln Ala Met Asp Asp Leu Met Leu Ser Pro Asp          35 40 45 Asp Ile Glu Gln Trp Phe Thr Glu Asp Pro Gly Pro Asp Glu Ala Pro      50 55 60 Arg Met Pro Glu Ala Pro Pro Ala Pro Pro Ala Pro Pro Ala Pro Pro  65 70 75 80 Thr Pro Ala Ala Pro Ala Pro Ala Pro Ser Trp Pro Leu Ser Ser Ser                  85 90 95 Val Ser Ser Gln Lys Thr Tyr Gln Gly Ser Tyr Gly Phe Arg Leu Gly             100 105 110 Phe Leu His Ser Gly Thr Ala Lys Ser Val Thr Cys Thr Tyr Ser Pro         115 120 125 Ala Leu Asn Lys Met Phe Cys Gln Leu Ala Lys Thr Cys Pro Val Gln     130 135 140 Leu Trp Val Asp Ser Thr Pro Pro Gly Thr Arg Val Arg Ala Met 145 150 155 160 Ala Ile Tyr Lys Gln Ser Gln His Met Thr Glu Val Val Arg Arg Cys                 165 170 175 Pro His His Glu Arg Cys Ser Asp Ser Asp Gly Leu Ala Pro Pro Gln             180 185 190 His Leu Ile Arg Val Glu Gly Asn Leu Arg Val Glu Tyr Leu Asp Asp         195 200 205 Arg Asn Thr Phe Arg His Ser Val Val Val Pro Tyr Glu Pro Pro Glu     210 215 220 Val Gly Ser Asp Cys Thr Thr Ile His Tyr Asn Tyr Met Cys Asn Ser 225 230 235 240 Ser Cys Met Gly Gly Met Asn Arg Arg Pro Ile Leu Thr Ile Ile Thr                 245 250 255 Leu Glu Asp Ser Ser Gly Asn Leu Leu Gly Arg Asn Ser Phe Glu Val             260 265 270 Arg Val Cys Ala Cys Pro Gly Arg Asp Arg Arg Thr Glu Glu Glu Asn         275 280 285 Leu Arg Lys Lys Gly Glu Pro His His Glu Leu Pro Pro Gly Ser Thr     290 295 300 Lys Arg Ala Leu Pro Asn Asn Thr Ser Ser Ser Pro Gln Pro Lys Lys 305 310 315 320 Lys Pro Leu Asp Gly Glu Tyr Phe Thr Leu Gln Ile Arg Gly Arg Glu                 325 330 335 Arg Phe Glu Met Phe Arg Glu Leu Asn Glu Ala Leu Glu Leu Lys Asp             340 345 350 Ala Gln Ala Gly Lys Glu Pro Gly Gly Ser Ser Ala His Ser Ser His         355 360 365 Leu Lys Ser Lys Lys Gly Gln Ser Thr Ser Arg His Lys Lys Leu Met     370 375 380 Phe Lys Thr Glu Gly Pro Asp Ser Asp 385 390 <210> 22 <211> 2843 <212> PRT <213> Homo sapiens_ APC amino acid <400> 22 Met Ala Ala Ala Ser Tyr Asp Gln Leu Leu Lys Gln Val Glu Ala Leu   1 5 10 15 Lys Met Glu Asn Ser Asn Leu Arg Gln Glu Leu Glu Asp Asn Ser Asn              20 25 30 His Leu Thr Lys Leu Glu Thr Glu Ala Ser Asn Met Lys Glu Val Leu          35 40 45 Lys Gln Leu Gln Gly Ser Ile Glu Asp Glu Ala Met Ala Ser Ser Gly      50 55 60 Gln Ile Asp Leu Leu Glu Arg Leu Lys Glu Leu Asn Leu Asp Ser Ser  65 70 75 80 Asn Phe Pro Gly Val Lys Leu Arg Ser Ser Met Ser Leu Arg Ser Tyr                  85 90 95 Gly Ser Arg Glu Gly Ser Val Ser Ser Ser Arg Gly Glu Cys Ser Pro             100 105 110 Val Pro Met Gly Ser Phe Pro Arg Arg Gly Phe Val Asn Gly Ser Arg         115 120 125 Glu Ser Thr Gly Tyr Leu Glu Glu Leu Glu Lys Glu Arg Ser Leu Leu     130 135 140 Leu Ala Asp Leu Asp Lys Glu Glu Lys Glu Lys Asp Trp Tyr Tyr Ala 145 150 155 160 Gln Leu Gln Asn Leu Thr Lys Arg Ile Asp Ser Leu Pro Leu Thr Glu                 165 170 175 Asn Phe Ser Leu Gln Thr Asp Met Thr Arg Arg Gln Leu Glu Tyr Glu             180 185 190 Ala Arg Gln Ile Arg Val Ala Met Glu Glu Gln Leu Gly Thr Cys Gln         195 200 205 Asp Met Glu Lys Arg Ala Gln Arg Arg Ile Ala Arg Ile Gln Gln Ile     210 215 220 Glu Lys Asp Ile Leu Arg Ile Arg Gln Leu Leu Gln Ser Gln Ala Thr 225 230 235 240 Glu Ala Glu Arg Ser Ser Gln Asn Lys His Glu Thr Gly Ser His Asp                 245 250 255 Ala Glu Arg Gln Asn Glu Gly Gln Gly Val Gly Glu Ile Asn Met Ala             260 265 270 Thr Ser Gly Asn Gly Gln Gly Ser Thr Thr Arg Met Asp His Glu Thr         275 280 285 Ala Ser Val Leu Ser Ser Ser Ser Thr His Ser Ala Pro Arg Arg Leu     290 295 300 Thr Ser His Leu Gly Thr Lys Val Glu Met Val Tyr Ser Leu Leu Ser 305 310 315 320 Met Leu Gly Thr His Asp Lys Asp Asp Met Ser Arg Thr Leu Leu Ala                 325 330 335 Met Ser Ser Ser Gln Asp Ser Cys Ile Ser Met Arg Gln Ser Gly Cys             340 345 350 Leu Pro Leu Leu Ile Gln Leu Leu His Gly Asn Asp Lys Asp Ser Val         355 360 365 Leu Leu Gly Asn Ser Arg Gly Ser Lys Glu Ala Arg Ala Arg Ala Ser     370 375 380 Ala Ala Leu His Asn Ile Ile His Ser Gln Pro Asp Asp Lys Arg Gly 385 390 395 400 Arg Arg Glu Ile Arg Val Leu His Leu Leu Glu Gln Ile Arg Ala Tyr                 405 410 415 Cys Glu Thr Cys Trp Glu Trp Gln Glu Ala His Glu Pro Gly Met Asp             420 425 430 Gln Asp Lys Asn Pro Met Pro Ala Pro Val Glu His Gln Ile Cys Pro         435 440 445 Ala Val Cys Val Leu Met Lys Leu Ser Phe Asp Glu Glu His Arg His     450 455 460 Ala Met Asn Glu Leu Gly Gly Leu Gln 465 470 475 480 Val Asp Cys Glu Met Tyr Gly Leu Thr Asn Asp His Tyr Ser Ile Thr                 485 490 495 Leu Arg Arg Tyr Ala Gly Met Ala Leu Thr Asn Leu Thr Phe Gly Asp             500 505 510 Val Ala Asn Lys Ala Thr Leu Cys Ser Met Lys Gly Cys Met Arg Ala         515 520 525 Leu Val Ala Gln Leu Lys Ser Glu Ser Glu Asp Leu Gln Gln Val Ile     530 535 540 Ala Ser Val Leu Arg Asn Leu Ser Trp Arg Ala Asp Val Asn Ser Lys 545 550 555 560 Lys Thr Leu Arg Glu Val Gly Ser Val Lys Ala Leu Met Glu Cys Ala                 565 570 575 Leu Glu Val Lys Lys Glu Ser Thr Leu Lys Ser Val Leu Ser Ala Leu             580 585 590 Trp Asn Leu Ser Ala His Cys Thr Glu Asn Lys Ala Asp Ile Cys Ala         595 600 605 Val Asp Gly Ala Leu Ala Phe Leu Val Gly Thr Leu Thr Tyr Arg Ser     610 615 620 Gln Thr Asn Thr Leu Ala Ile Ile Glu Ser Gly Gly Gly Ile Leu Arg 625 630 635 640 Asn Val Ser Ser Leu Ile Ala Thr Asn Glu Asp His Arg Gln Ile Leu                 645 650 655 Arg Glu Asn Asn Cys Leu Gln Thr Leu Leu Gln His Leu Lys Ser His             660 665 670 Ser Leu Thr Ile Val Ser Asn Ala Cys Gly Thr Leu Trp Asn Leu Ser         675 680 685 Ala Arg Asn Pro Lys Asp Gln Glu Ala Leu Trp Asp Met Gly Ala Val     690 695 700 Ser Met Leu Lys Asn Leu Ile His Ser Lys His Lys Met Ile Ala Met 705 710 715 720 Gly Ser Ala Ala Ala Leu Arg Asn Leu Met Ala Asn Arg Pro Ala Lys                 725 730 735 Tyr Lys Asp Ala Asn Ile Met Ser Pro Gly Ser Ser Leu Pro Ser Leu             740 745 750 His Val Arg Lys Gln Lys Ala Leu Glu Ala Glu Leu Asp Ala Gln His         755 760 765 Leu Ser Glu Thr Phe Asp Asn Ile Asp Asn Leu Ser Pro Lys Ala Ser     770 775 780 His Arg Ser Lys Gln Arg His Lys Gln Ser Leu Tyr Gly Asp Tyr Val 785 790 795 800 Phe Asp Thr Asn Arg His Asp Asp Asn Arg Ser Asp Asn Phe Asn Thr                 805 810 815 Gly Asn Met Thr Val Leu Ser Pro Tyr Leu Asn Thr Thr Val Leu Pro             820 825 830 Ser Ser Ser Ser Ser Glu Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser         835 840 845 Asp Arg Ser Leu Glu Arg Glu Arg Gly Ile Gly Leu Gly Asn Tyr His     850 855 860 Pro Ala Thr Glu Asn Pro Gly Thr Ser Ser Lys Arg Gly Leu Gln Ile 865 870 875 880 Ser Thr Thr Ala Ala Gln Ile Ala Lys Val Met Glu Glu Val Ser Ala                 885 890 895 Ile His Thr Ser Gln Glu Asp Arg Ser Ser Gly Ser Thr Thr Glu Leu             900 905 910 His Cys Val Thr Asp Glu Arg Asn Ala Leu Arg Arg Ser Ser Ala Ala         915 920 925 His Thr His Ser Asn Thr Tyr Asn Phe Thr Lys Ser Glu Asn Ser Asn     930 935 940 Arg Thr Cys Ser Met Pro Tyr Ala Lys Leu Glu Tyr Lys Arg Ser Ser 945 950 955 960 Asn Asp Ser Leu Asn Ser Val Ser Ser Ser Asp Gly Tyr Gly Lys Arg                 965 970 975 Gly Gln Met Lys Pro Ser Ile Glu Ser Tyr Ser Glu Asp Asp Glu Ser             980 985 990 Lys Phe Cys Ser Tyr Gly Gln Tyr Pro Ala Asp Leu Ala His Lys Ile         995 1000 1005 His Ser Ala Asn His Met Asp Asp Asn Asp Gly Glu Leu Asp Thr Pro    1010 1015 1020 Ile Asn Tyr Ser Leu Lys Tyr Ser Asp Glu Gln Leu Asn Ser Gly Arg 1025 1030 1035 1040 Gln Ser Ser Gln Asn Glu Arg Trp Ala Arg Pro Lys His Ile Ile                1045 1050 1055 Glu Asp Glu Ile Lys Gln Ser Glu Gln Arg Gln Ser Arg Asn Gln Ser            1060 1065 1070 Thr Thr Tyr Pro Val Tyr Thr Glu Ser Thr Asp Asp Lys His Leu Lys        1075 1080 1085 Phe Gln Pro His Phe Gly Gln Gin Glu Cys Val Ser Pro Tyr Arg Ser    1090 1095 1100 Arg Gly Ala Asn Gly Ser Glu Thr Asn Arg Val Gly Ser Asn His Gly 1105 1110 1115 1120 Ile Asn Gln Asn Val Ser Gln Ser Leu Cys Gln Glu Asp Asp Tyr Glu                1125 1130 1135 Asp Asp Lys Pro Thr Asn Tyr Ser Glu Arg Tyr Ser Glu Glu Glu Gln            1140 1145 1150 His Glu Glu Glu Glu Arg Pro Thr Asn Tyr Ser Ile Lys Tyr Asn Glu        1155 1160 1165 Glu Lys Arg His Val Asp Gln Pro Ile Asp Tyr Ser Leu Lys Tyr Ala    1170 1175 1180 Thr Asp Ile Pro Ser Ser Gln Lys Gln Ser Ser Phe Ser Ser Ser Ser Ser Ser 1185 1190 1195 1200 Ser Ser Gly Gln Ser Ser Lys Thr Glu His Met Ser Ser Ser Glu                1205 1210 1215 Asn Thr Ser Thr Ser Ser Asn Ala Lys Arg Gln Asn Gln Leu His            1220 1225 1230 Pro Ser Ser Ala Gln Ser Ser Ser Gly Gln Pro Gln Lys Ala Ala Thr        1235 1240 1245 Cys Lys Val Ser Ser Ile Asn Gln Glu Thr Ile Gln Thr Tyr Cys Val    1250 1255 1260 Glu Asp Thr Pro Ile Cys Phe Ser Ser Cys Ser Ser Leu Ser Ser Leu 1265 1270 1275 1280 Ser Ser Ala Glu Asp Glu Ile Gly Cys Asn Gln Thr Thr Gln Glu Ala                1285 1290 1295 Asp Ser Ala Asn Thr Leu Gln Ile Ala Glu Ile Lys Glu Lys Ile Gly            1300 1305 1310 Thr Arg Ser Ala Glu Asp Pro Val Ser Glu Val Pro Ala Val Ser Gln        1315 1320 1325 His Pro Arg Thr Lys Ser Ser Arg Leu Gln Gly Ser Ser Leu Ser Ser    1330 1335 1340 Glu Ser Ala Arg His Lys Ala Val Glu Phe Ser Ser Gly Ala Lys Ser 1345 1350 1355 1360 Pro Ser Lys Ser Gly Ala Gln Thr Pro Lys Ser Pro Pro Glu His Tyr                1365 1370 1375 Val Gln Glu Thr Pro Leu Met Phe Ser Arg Cys Thr Ser Val Ser Ser            1380 1385 1390 Leu Asp Ser Phe Glu Ser Ser Ser Ile Ala Ser Ser Val Gln Ser Glu        1395 1400 1405 Pro Cys Ser Gly Met Val Ser Gly Ile Ile Ser Pro Ser Asp Leu Pro    1410 1415 1420 Asp Ser Pro Gly Gln Thr Met Pro Pro Ser Arg Ser Lys Thr Pro Pro 1425 1430 1435 1440 Pro Pro Pro Gln Thr Ala Gln Thr Lys Arg Glu Val Pro Lys Asn Lys                1445 1450 1455 Ala Pro Thr Ala Glu Lys Arg Glu Ser Gly Pro Lys Gln Ala Ala Val            1460 1465 1470 Asn Ala Ala Val Gln Arg Val Gln Val Leu Pro Asp Ala Asp Thr Leu        1475 1480 1485 Leu His Phe Ala Thr Glu Ser Thr Pro Asp Gly Phe Ser Cys Ser Ser    1490 1495 1500 Ser Leu Ser Ala Leu Ser Leu Asp Glu Pro Phe Ile Gln Lys Asp Val 1505 1510 1515 1520 Glu Leu Arg Ile Met Pro Pro Val Glu Glu Asn Asp Asn Gly Asn Glu                1525 1530 1535 Thr Glu Ser Glu Gln Pro Lys Glu Ser Asn Glu Asn Gln Glu Lys Glu            1540 1545 1550 Ala Glu Lys Thr Ile Asp Ser Glu Lys Asp Leu Leu Asp Asp Ser Asp        1555 1560 1565 Asp Asp Asp Ile Glu Ile Leu Glu Glu Cys Ile Ser Ser Ala Met Pro    1570 1575 1580 Thr Lys Ser Ser Arg Lys Ala Lys Lys Pro Ala Gln Thr Ala Ser Lys 1585 1590 1595 1600 Leu Pro Pro Val Ala Arg Lys Pro Ser Gln Leu Pro Val Tyr Lys                1605 1610 1615 Leu Leu Pro Ser Gln Asn Arg Leu Gln Pro Gln Lys His Val Ser Phe            1620 1625 1630 Thr Pro Gly Asp Asp Met Pro Arg Val Tyr Cys Val Glu Gly Thr Pro        1635 1640 1645 Ile Asn Phe Ser Thr Ala Thr Ser Leu Ser Asp Leu Thr Ile Glu Ser    1650 1655 1660 Pro Pro Asn Glu Leu Ala Ala Gly Glu Gly Val Arg Gly Gly Ala Gln 1665 1670 1675 1680 Ser Gly Glu Phe Glu Lys Arg Asp Thr Ile Pro Thr Glu Gly Arg Ser                1685 1690 1695 Thr Asp Glu Gly Gly Gly Lys Thr Ser Ser Val Thr Ile Pro Glu            1700 1705 1710 Leu Asp Asp Asn Lys Ala Glu Glu Gly Asp Ile Leu Ala Glu Cys Ile        1715 1720 1725 Asn Ser Ala Met Pro Lys Gly Lys Ser His Lys Pro Phe Arg Val Lys    1730 1735 1740 Lys Ile Met Asp Gln Val Gln Gln Ala Ser Ala Ser Ser Ser Ala Pro 1745 1750 1755 1760 Asn Lys Asn Gln Leu Asp Gly Lys Lys Lys Lys Pro Thr Ser Pro Val                1765 1770 1775 Lys Pro Ile Pro Gln Asn Thr Glu Tyr Arg Thr Arg Val Arg Lys Asn            1780 1785 1790 Ala Asp Ser Lys Asn Asn Leu Asn Ala Glu Arg Val Phe Ser Asp Asn        1795 1800 1805 Lys Asp Ser Lys Lys Gln Asn Leu Lys Asn Asn Ser Lys Val Phe Asn    1810 1815 1820 Asp Lys Leu Pro Asn Asn Glu Asp Arg Val Gly Ser Phe Ala Phe 1825 1830 1835 1840 Asp Ser Pro His His Tyr Thr Pro Ile Glu Gly Thr Pro Tyr Cys Phe                1845 1850 1855 Ser Arg Asn Asp Ser Leu Ser Ser Leu Asp Phe Asp Asp Asp Asp Val            1860 1865 1870 Asp Leu Ser Arg Glu Lys Ala Glu Leu Arg Lys Ala Lys Glu Asn Lys        1875 1880 1885 Glu Ser Glu Ala Lys Val Thr Ser His Thr Glu Leu Thr Ser Asn Gln    1890 1895 1900 Gln Ser Ala Asn Lys Thr Gln Ala Ile Ala Lys Gln Pro Ile Asn Arg 1905 1910 1915 1920 Gly Gln Pro Lys Pro Ile Leu Gln Lys Gln Ser Thr Phe Pro Gln Ser                1925 1930 1935 Ser Lys Asp Ile Pro Asp Arg Gly Ala Ala Thr Asp Glu Lys Leu Gln            1940 1945 1950 Asn Phe Ala Ile Glu Asn Thr Pro Val Cys Phe Ser His Asn Ser Ser        1955 1960 1965 Leu Ser Ser Leu Ser Asp Ile Asp Gln Glu Asn Asn Asn Lys Glu Asn    1970 1975 1980 Glu Pro Ile Lys Glu Thr Glu Pro Pro Asp Ser Gln Gly Glu Pro Ser 1985 1990 1995 2000 Lys Pro Gln Ala Ser Gly Tyr Ala Pro Lys Ser Phe His Val Glu Asp                2005 2010 2015 Thr Pro Val Cys Phe Ser Arg Asn Ser Ser Leu Ser Ser Leu Ser Ile            2020 2025 2030 Asp Ser Glu Asp Asp Leu Leu Gln Glu Cys Ile Ser Ser Ala Met Pro        2035 2040 2045 Lys Lys Lys Lys Pro Ser Arg Leu Lys Gly Asp Asn Glu Lys His Ser    2050 2055 2060 Pro Arg Asn Met Gly Gly Ile Leu Gly Glu Asp Leu Thr Leu Asp Leu 2065 2070 2075 2080 Lys Asp Ile Gln Arg Pro Asp Ser Glu His Gly Leu Ser Pro Asp Ser                2085 2090 2095 Glu Asn Phe Asp Trp Lys Ala Ile Gln Glu Gly Ala Asn Ser Ile Val            2100 2105 2110 Ser Ser Leu His Gln Ala Ala Ala Ala Cys Leu Ser Arg Gln Ala        2115 2120 2125 Ser Ser As Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Ser Soy    2130 2135 2140 Gly Ser Pro Phe His Leu Thr Pro Asp Gln Glu Glu Lys Pro Phe Thr 2145 2150 2155 2160 Ser Asn Lys Gly Pro Arg Ile Leu Lys Pro Gly Glu Lys Ser Thr Leu                2165 2170 2175 Glu Thr Lys Lys Ile Glu Ser Glu Ser Lys Gly Ile Lys Gly Gly Lys            2180 2185 2190 Lys Val Tyr Lys Ser Leu Ile Thr Gly Lys Val Arg Ser Ser Ser Ser        2195 2200 2205 Ile Ser Gly Gln Met Lys Gln Pro Leu Gln Ala Asn Met Pro Ser Ile    2210 2215 2220 Ser Arg Gly Arg Thr Met Ile His Ile Pro Gly Val Arg Asn Ser Ser 2225 2230 2235 2240 Ser Ser Thr Ser Pro Val Ser Lys Lys Gly Pro Pro Leu Lys Thr Pro                2245 2250 2255 Ala Ser Lys Ser Ser Ser Glu Gly Gln Thr Ala Thr Ser Ser Arg            2260 2265 2270 Gly Ala Lys Pro Ser Val Lys Ser Glu Leu Ser Pro Val Ala Arg Gln        2275 2280 2285 Thr Ser Gln Ile Gly Gly Ser Ser Lys Ala Pro Ser Ser Ser Ser Ser Ser    2290 2295 2300 Arg Asp Ser Thr Pro Ser Arg Pro Ala Gln Gln Pro Leu Ser Arg Pro 2305 2310 2315 2320 Ile Gln Ser Pro Gly Arg Asn Ser Ile Ser Pro Gly Arg Asn Gly Ile                2325 2330 2335 Ser Pro Pro Asn Lys Leu Ser Gln Leu Pro Arg Thr Ser Ser Ser Ser            2340 2345 2350 Thr Ala Ser Thr Lys Ser Ser Gly Ser Gly Lys Met Ser Ser Thyr Ser        2355 2360 2365 Pro Gly Arg Gln Met Ser Gln Gln Asn Leu Thr Lys Gln Thr Gly Leu    2370 2375 2380 Ser Lys Asn Ala Ser Ser Ile Pro Arg Ser Glu Ser Ala Ser Lys Gly 2385 2390 2395 2400 Leu Asn Gln Met Asn Asn Gly Asn Gly Ala Asn Lys Lys Val Glu Leu                2405 2410 2415 Ser Arg Met Ser Ser Thr Lys Ser Ser Gly Ser Glu Ser Asp Arg Ser            2420 2425 2430 Glu Arg Pro Val Leu Val Arg Gln Ser Thr Phe Ile Lys Glu Ala Pro        2435 2440 2445 Ser Pro Thr Leu Arg Arg Lys Leu Glu Glu Ser Ala Ser Phe Glu Ser    2450 2455 2460 Leu Ser Pro Ser Ser Arg Pro Ala Ser Pro Thr Arg Ser Gln Ala Gln 2465 2470 2475 2480 Thr Pro Val Leu Ser Pro Ser Leu Pro Asp Met Ser Leu Ser Thr His                2485 2490 2495 Ser Ser Val Gln Ala Gly Gly Trp Arg Lys Leu Pro Pro Asn Leu Ser            2500 2505 2510 Pro Thr Ile Glu Tyr Asn Asp Gly Arg Pro Ala Lys Arg His Asp Ile        2515 2520 2525 Ala Arg Ser His Ser Glu Ser Pro Ser Arg Leu Pro Ile Asn Arg Ser    2530 2535 2540 Gly Thr Trp Lys Arg Glu His Ser Lys His Ser Ser Ser Leu Pro Arg 2545 2550 2555 2560 Val Ser Thr Trp Arg Arg Thr Gly Ser Ser Ser Ser Le Le Ser Ala                2565 2570 2575 Ser Ser Glu Ser Ser Glu Lys Ala Lys Ser Glu Asp Glu Lys His Val            2580 2585 2590 Asn Ser Ile Ser Gly Thr Lys Gln Ser Lys Glu Asn GIn Val Ser Ala        2595 2600 2605 Lys Gly Thr Trp Arg Lys Ile Lys Glu Asn Glu Phe Ser Pro Thr Asn    2610 2615 2620 Ser Thr Ser Gln Thr Val Ser Ser Gly Ala Thr Asn Gly Ala Glu Ser 2625 2630 2635 2640 Lys Thr Leu Ile Tyr Gln Met Ala Pro Ala Val Ser Lys Thr Glu Asp                2645 2650 2655 Val Trp Val Arg Ile Glu Asp Cys Pro Ile Asn Asn Pro Arg Ser Gly            2660 2665 2670 Arg Ser Pro Thr Gly Asn Thr Pro Pro Val Ile Asp Ser Val Ser Glu        2675 2680 2685 Lys Ala Asn Pro Asn Ile Lys Asp Ser Lys Asp Asn Gln Ala Lys Gln    2690 2695 2700 Asn Val Gly Asn Gly Ser Val Pro Met Met Thr Val Gly Leu Glu Asn 2705 2710 2715 2720 Arg Leu Asn Ser Phe Ile Gln Val Asp Ala Pro Asp Gln Lys Gly Thr                2725 2730 2735 Glu Ile Lys Pro Gly Gln Asn Asn Pro Val Val Val Ser Glu Thr Asn            2740 2745 2750 Glu Ser Ser Val Glu Arg Thr Pro Ser Ser Ser Ser Ser Ser        2755 2760 2765 Lys His Ser Ser Ser Gly Thr Val Ala Ala Arg Val Thr Pro Phe    2770 2775 2780 Asn Tyr Asn Pro Ser Pro Arg Lys Ser Ser Ala Asp Ser Thr Ser Ala 2785 2790 2795 2800 Arg Pro Ser Gln Ile Pro Thr Pro Val Asn Asn Asn Thr Lys Lys Arg                2805 2810 2815 Asp Ser Lys Thr Asp Ser Thr Glu Ser Ser Gly Thr Gln Ser Pro Lys            2820 2825 2830 Arg His Ser Gly Ser Tyr Leu Val Thr Ser Val        2835 2840 <210> 23 <211> 1068 <212> PRT <213> Homo sapiens_ PIK3CA amino acid <400> 23 Met Pro Pro Arg Ser Ser Gly Glu Leu Trp Gly Ile His Leu Met   1 5 10 15 Pro Pro Arg Ile Leu Val Glu Cys Leu Leu Pro Asn Gly Met Ile Val              20 25 30 Thr Leu Glu Cys Leu Arg Glu Ala Thr Leu Val Thr Ile Lys His Glu          35 40 45 Leu Phe Lys Glu Ala Arg Lys Tyr Pro Leu His Gln Leu Leu Gln Asp      50 55 60 Glu Ser Ser Tyr Ile Phe Val Ser Val Thr Gln Glu Ala Glu Arg Glu  65 70 75 80 Glu Phe Phe Asp Glu Thr Arg Arg Leu Cys Asp Leu Arg Leu Phe Gln                  85 90 95 Pro Phe Leu Lys Val Ile Glu Pro Val Gly Asn Arg Glu Glu Lys Ile             100 105 110 Leu Asn Arg Glu Ile Gly Phe Ala Ile Gly Met Pro Val Cys Glu Phe         115 120 125 Asp Met Val Lys Asp Pro Glu Val Gln Asp Phe Arg Arg Asn Ile Leu     130 135 140 Asn Val Cys Lys Glu Ala Val Asp Leu Arg Asp Leu Asn Ser Pro His 145 150 155 160 Ser Arg Ala Met Tyr Val Tyr Pro Pro His Val Glu Ser Ser Pro Glu                 165 170 175 Leu Pro Lys His Ile Tyr Asn Lys Leu Asp Arg Gly Gln Ile Ile Val             180 185 190 Val Ile Trp Val Ile Val Ser Pro Asn Asn Asp Lys Gln Lys Tyr Thr         195 200 205 Leu Lys Ile Asn His Asp Cys Val Pro Glu Gln Val Ile Ala Glu Ala     210 215 220 Ile Arg Lys Lys Thr Arg Ser Met Leu Leu Ser Ser Glu Gln Leu Lys 225 230 235 240 Leu Cys Val Leu Glu Tyr Gln Gly Lys Tyr Ile Leu Lys Val Cys Gly                 245 250 255 Cys Asp Glu Tyr Phe Leu Glu Lys Tyr Pro Leu Ser Gln Tyr Lys Tyr             260 265 270 Ile Arg Ser Cys Ile Met Leu Gly Arg Met Pro Asn Leu Lys Met Met         275 280 285 Ala Lys Glu Ser Leu Tyr Ser Gln Leu Pro Met Asp Cys Phe Thr Met     290 295 300 Pro Ser Tyr Ser Arg Arg Ser Ser Thr Ala Thr Pro Tyr Met Asn Gly 305 310 315 320 Glu Thr Ser Thr Lys Ser Leu Trp Val Ile Asn Arg Ala Leu Arg Ile                 325 330 335 Lys Ile Leu Cys Ala Thr Tyr Val Asn Leu Asn Ile Arg Asp Ile Asp             340 345 350 Lys Ile Tyr Val Arg Thr Gly Ile Tyr His Gly Gly Glu Pro Leu Cys         355 360 365 Asp Asn Val Asn Thr Gln Arg Val Pro Cys Ser Asn Pro Arg Trp Asn     370 375 380 Glu Trp Leu Asn Tyr Asp Ile Tyr Ile Pro Asp Leu Pro Arg Ala Ala 385 390 395 400 Arg Leu Cys Leu Ser Ile Cys Ser Val Lys Gly Arg Lys Gly Ala Lys                 405 410 415 Glu Glu His Cys Pro Leu Ala Trp Gly Asn Ile Asn Leu Phe Asp Tyr             420 425 430 Thr Asp Thr Leu Val Ser Gly Lys Met Ala Leu Asn Leu Trp Pro Val         435 440 445 Pro His Gly Leu Glu Asp Leu Leu Asn Pro Ile Gly Val Thr Gly Ser     450 455 460 Asn Pro Asn Lys Glu Thr Pro Cys Leu Glu Leu Glu Phe Asp Trp Phe 465 470 475 480 Ser Ser Val Val Lys Phe Pro Asp Met Ser Val Ile Glu Glu His Ala                 485 490 495 Asn Trp Ser Val Ser Arg Glu Ala Gly Phe Ser Tyr Ser His Ala Gly             500 505 510 Leu Ser Asn Arg Leu Ala Arg Asp Asn Glu Leu Arg Glu Asn Asp Lys         515 520 525 Glu Gln Leu Lys Ala Ile Ser Thr Arg Asp Pro Leu Ser Glu Ile Thr     530 535 540 Glu Gln Glu Lys Asp Phe Leu Trp Ser His Arg His Tyr Cys Val Thr 545 550 555 560 Ile Pro Glu Ile Leu Pro Lys Leu Leu Leu Ser Val Lys Trp Asn Ser                 565 570 575 Arg Asp Glu Val Ala Gln Met Tyr Cys Leu Val Lys Asp Trp Pro Pro             580 585 590 Ile Lys Pro Glu Gln Ala Met Glu Leu Leu Asp Cys Asn Tyr Pro Asp         595 600 605 Pro Met Val Arg Gly Phe Ala Val Arg Cys Leu Glu Lys Tyr Leu Thr     610 615 620 Asp Asp Lys Leu Ser Gln Tyr Leu Ile Gln Leu Val Gln Val Leu Lys 625 630 635 640 Tyr Glu Gln Tyr Leu Asp Asn Leu Leu Val Arg Phe Leu Leu Lys Lys                 645 650 655 Ala Leu Thr Asn Gln Arg Ile Gly His Phe Phe Phe Trp His Leu Lys             660 665 670 Ser Glu Met His Asn Lys Thr Val Ser Gln Arg Phe Gly Leu Leu Leu         675 680 685 Glu Ser Tyr Cys Arg Ala Cys Gly Met Tyr Leu Lys His Leu Asn Arg     690 695 700 Gln Val Glu Ala Met Glu Lys Leu Ile Asn Leu Thr Asp Ile Leu Lys 705 710 715 720 Gln Glu Arg Lys Asp Glu Thr Gln Lys Val Gln Met Lys Phe Leu Val                 725 730 735 Glu Gln Met Arg Arg Pro Asp Phe Met Asp Ala Leu Gln Gly Leu Leu             740 745 750 Ser Pro Leu Asn Pro Ala His Gln Leu Gly Asn Leu Arg Leu Lys Glu         755 760 765 Cys Arg Ile Met Ser Ser Ala Lys Arg Pro Leu Trp Leu Asn Trp Glu     770 775 780 Asn Pro Asp Ile Met Ser Glu Leu Leu Phe Gln Asn Asn Glu Ile Ile 785 790 795 800 Phe Lys Asn Gly Asp Asp Leu Arg Gln Asp Met Leu Thr Leu Gln Ile                 805 810 815 Ile Arg Ile Met Glu Asn Ile Trp Gln Asn Gln Gly Leu Asp Leu Arg             820 825 830 Met Leu Pro Tyr Gly Cys Leu Ser Ile Gly Asp Cys Val Gly Leu Ile         835 840 845 Glu Val Val Arg Asn Ser His Thr Ile Met Gln Ile Gln Cys Lys Gly     850 855 860 Gly Leu Lys Gly Ala Leu Gln Phe Asn Ser His Thr Leu His Gln Trp 865 870 875 880 Leu Lys Asp Lys Asn Lys Gly Glu Ile Tyr Asp Ala Ala Ile Asp Leu                 885 890 895 Phe Thr Arg Ser Cys Ala Gly Tyr Cys Val Ala Thr Phe Ile Leu Gly             900 905 910 Ile Gly Asp Arg His Asn Ser Asn Ile Met Val Lys Asp Asp Gly Gln         915 920 925 Leu Phe His Ile Asp Phe Gly His Phe Leu Asp His Lys Lys Lys Lys     930 935 940 Phe Gly Tyr Lys Arg Glu Arg Val Pro Phe Val Leu Thr Gln Asp Phe 945 950 955 960 Leu Ile Val Ile Ser Lys Gly Ala Gln Glu Cys Thr Lys Thr Arg Glu                 965 970 975 Phe Glu Arg Phe Gln Glu Met Cys Tyr Lys Ala Tyr Leu Ala Ile Arg             980 985 990 Gln His Ala Asn Leu Phe Ile Asn Leu Phe Ser Met Met Leu Gly Ser         995 1000 1005 Gly Met Pro Glu Leu Gln Ser Phe Asp Asp Ile Ala Tyr Ile Arg Lys    1010 1015 1020 Thr Leu Ala Leu Asp Lys Thr Glu Gln Glu Ala Leu Glu Tyr Phe Met 1025 1030 1035 1040 Lys Gln Met Asn Asp Ala His His Gly Gly Trp Thr Thr Lys Met Asp                1045 1050 1055 Trp Ile Phe His Thr Ile Lys Gln His Ala Leu Asn            1060 1065 <210> 24 <211> 188 <212> PRT <213> Homo sapiens_ KRAS amino acid <400> 24 Met Thr Glu Tyr Lys Leu Val Val Gly Aly Gly Gly Val Gly Lys   1 5 10 15 Ser Ala Leu Thr Ile Gln Leu Ile Gln Asn His Phe Val Asp Glu Tyr              20 25 30 Asp Pro Thr Ile Glu Asp Ser Tyr Arg Lys Gln Val Val Ile Asp Gly          35 40 45 Glu Thr Cys Leu Leu Asp Ile Leu Asp Thr Ala Gly Gln Glu Glu Tyr      50 55 60 Ser Ala Met Arg Asp Gln Tyr Met Arg Thr Gly Glu Gly Phe Leu Cys  65 70 75 80 Val Phe Ala Ile Asn Asn Thr Lys Ser Phe Glu Asp Ile His His Tyr                  85 90 95 Arg Glu Gln Ile Lys Arg Val Lys Asp Ser Glu Asp Val Pro Met Val             100 105 110 Leu Val Gly Asn Lys Cys Asp Leu Pro Ser Arg Thr Val Asp Thr Lys         115 120 125 Gln Ala Gln Asp Leu Ala Arg Ser Tyr Gly Ile Pro Phe Ile Glu Thr     130 135 140 Ser Ala Lys Thr Arg Gln Gly Val Asp Asp Ala Phe Tyr Thr Leu Val 145 150 155 160 Arg Glu Ile Arg Lys His Lys Glu Lys Met Ser Lys Asp Gly Lys Lys                 165 170 175 Lys Lys Lys Lys Ser Lys Thr Lys Cys Val Ile Met             180 185 <210> 25 <211> 787 <212> PRT <213> Homo sapiens_ SMO amino acid <400> 25 Met Ala Ala Ala Arg Pro Ala Arg Gly Pro Glu Leu Pro Leu Leu Gly   1 5 10 15 Leu Leu Leu Leu Leu Leu Gly Asp Pro Gly Arg Gly Ala Ala Ser              20 25 30 Ser Gly Asn Ala Thr Gly Pro Gly Pro Arg Ser Ala Gly Gly Ser Ala          35 40 45 Arg Arg Ser Ala Ala Val Thr Gly Pro Pro Pro Pro Leu Ser His Cys      50 55 60 Gly Arg Ala Ala Pro Cys Glu Pro Leu Arg Tyr Asn Val Cys Leu Gly  65 70 75 80 Ser Val Leu Pro Tyr Gly Ala Thr Ser Thr Leu Leu Ala Gly Asp Ser                  85 90 95 Asp Ser Gln Glu Glu Ala His Gly Lys Leu Val Leu Trp Ser Gly Leu             100 105 110 Arg Asn Ala Pro Arg Cys Trp Ala Val Ile Gln Pro Leu Leu Cys Ala         115 120 125 Val Tyr Met Pro Lys Cys Glu Asn Asp Arg Val Glu Leu Pro Ser Arg     130 135 140 Thr Leu Cys Gln Ala Thr Arg Gly Pro Cys Ala Ile Val Glu Arg Glu 145 150 155 160 Arg Gly Trp Pro Asp Phe Leu Arg Cys Thr Pro Asp Arg Phe Pro Glu                 165 170 175 Gly Cys Thr Asn Glu Val Gln Asn Ile Lys Phe Asn Ser Ser Gly Gln             180 185 190 Cys Glu Val Pro Leu Val Arg Thr Asp Asn Pro Lys Ser Trp Tyr Glu         195 200 205 Asp Val Glu Gly Cys Gly Ile Gln Cys Gln Asn Pro Leu Phe Thr Glu     210 215 220 Ala Glu His Gln Asp Met His Ser Tyr Ile Ala Ala Phe Gly Ala Val 225 230 235 240 Thr Gly Leu Cys Thr Leu Phe Thr Leu Ala Thr Phe Val Ala Asp Trp                 245 250 255 Arg Asn Ser Asn Arg Tyr Pro Ala Val Ile Leu Phe Tyr Val Asn Ala             260 265 270 Cys Phe Phe Val Gly Ser Ile Gly Trp Leu Ala Gln Phe Met Asp Gly         275 280 285 Ala Arg Arg Glu Ile Val Cys Arg Ala Asp Gly Thr Met Arg Leu Gly     290 295 300 Glu Pro Thr Ser Asn Glu Thr Leu Ser Cys Val Ile Ile Phe Val Ile 305 310 315 320 Val Tyr Tyr Ala Leu Met Ala Gly Val Val Trp Phe Val Val Leu Thr                 325 330 335 Tyr Ala Trp His Thr Ser Phe Lys Ala Leu Gly Thr Thr Tyr Gln Pro             340 345 350 Leu Ser Gly Lys Thr Ser Tyr Phe His Leu Leu Thr Trp Ser Leu Pro         355 360 365 Phe Val Leu Thr Val Ala Ile Leu Ala Val Ala Gln Val Asp Gly Asp     370 375 380 Ser Val Ser Gly Ile Cys Phe Val Gly Tyr Lys Asn Tyr Arg Tyr Arg 385 390 395 400 Ala Gly Phe Val Leu Ala Pro Ile Gly Leu Val Leu Ile Val Gly Gly                 405 410 415 Tyr Phe Leu Ile Arg Gly Val Met Thr Leu Phe Ser Ile Lys Ser Asn             420 425 430 His Pro Gly Leu Leu Ser Glu Lys Ala Ala Ser Lys Ile Asn Glu Thr         435 440 445 Met Leu Arg Leu Gly Ile Phe Gly Phe Leu Ala Phe Gly Phe Val Leu     450 455 460 Ile Thr Phe Ser Cys His Phe Tyr Asp Phe Phe Asn Gln Ala Glu Trp 465 470 475 480 Glu Arg Ser Phe Arg Asp Tyr Val Leu Cys Gln Ala Asn Val Thr Ile                 485 490 495 Gly Leu Pro Thr Lys Gln Pro Ile Pro Asp Cys Glu Ile Lys Asn Arg             500 505 510 Pro Ser Leu Leu Val Glu Lys Ile Asn Leu Phe Ala Met Phe Gly Thr         515 520 525 Gly Ile Ala Met Ser Thr Trp Val Trp Thr Lys Ala Thr Leu Leu Ile     530 535 540 Trp Arg Arg Thr Trp Cys Arg Leu Thr Gly Gln Ser Asp Asp Glu Pro 545 550 555 560 Lys Arg Ile Lys Lys Ser Lys Met Ile Ala Lys Ala Phe Ser Lys Arg                 565 570 575 His Glu Leu Leu Gln Asn Pro Gly Gln Glu Leu Ser Phe Ser Met His             580 585 590 Thr Val Ser His Asp Gly Pro Val Ala Gly Leu Ala Phe Asp Leu Asn         595 600 605 Glu Pro Ser Ala Asp Val Ser Ser Ala Trp Ala Gln His Val Thr Lys     610 615 620 Met Val Ala Arg Arg Gly Ala Ile Leu Pro Gln Asp Ile Ser Val Thr 625 630 635 640 Pro Val Ala Thr Pro Val Pro Pro Glu Glu Gln Ala Asn Leu Trp Leu                 645 650 655 Val Glu Ala Glu Ile Ser Pro Glu Leu Gln Lys Arg Leu Gly Arg Lys             660 665 670 Lys Lys Arg Arg Lys Arg Lys Lys Glu Val Cys Pro Leu Ala Pro Pro         675 680 685 Pro Glu Leu His Pro Pro Ala Pro Ala Pro Ser Thr Ile Pro Arg Leu     690 695 700 Pro Gln Leu Pro Arg Gln Lys Cys Leu Val Ala Gly Ala Trp Gly 705 710 715 720 Ala Gly Asp Ser Cys Arg Gln Gly Ala Trp Thr Leu Val Ser Asn Pro                 725 730 735 Phe Cys Pro Glu Pro Ser Pro Pro Gln Asp Pro Phe Leu Pro Ser Ala             740 745 750 Pro Ala Pro Val Ala Trp Ala His Gly Arg Arg Gln Gly Leu Gly Pro         755 760 765 Ile His Ser Arg Thr Asn Leu Met Asp Thr Glu Leu Met Asp Ala Asp     770 775 780 Ser Asp Phe 785 <210> 26 <211> 433 <212> PRT <213> Homo sapiens STK11 amino acid <400> 26 Met Glu Val Val Asp Pro Gln Gln Leu Gly Met Phe Thr Glu Gly Glu   1 5 10 15 Leu Met Ser Val Gly Met Asp Thr Phe Ile His Arg Ile Asp Ser Thr              20 25 30 Glu Val Ile Tyr Gln Pro Arg Arg Lys Arg Ala Lys Leu Ile Gly Lys          35 40 45 Tyr Leu Met Gly Asp Leu Leu Gly Glu Gly Ser Tyr Gly Lys Val Lys      50 55 60 Glu Val Leu Asp Ser Glu Thr Leu Cys Arg Arg Ala Val Lys Ile Leu  65 70 75 80 Lys Lys Lys Lys Leu Arg Arg Ile Pro Asn Gly Glu Ala Asn Val Lys                  85 90 95 Lys Glu Ile Gln Leu Leu Arg Arg Leu Arg His Lys Asn Val Ile Gln             100 105 110 Leu Val Asp Val Leu Tyr Asn Glu Glu Lys Gln Lys Met Tyr Met Val         115 120 125 Met Glu Tyr Cys Val Cys Gly Met Gln Glu Met Leu Asp Ser Val Pro     130 135 140 Glu Lys Arg Phe Pro Val Cys Gln Ala His Gly Tyr Phe Cys Gln Leu 145 150 155 160 Ile Asp Gly Leu Glu Tyr Leu His Ser Gln Gly Ile Val His Lys Asp                 165 170 175 Ile Lys Pro Gly Asn Leu Leu Leu Thr Thr Gly Gly Thr Leu Lys Ile             180 185 190 Ser Asp Leu Gly Val Ala Glu Ala Leu His Pro Phe Ala Ala Asp Asp         195 200 205 Thr Cys Arg Thr Ser Gln Gly Ser Pro Ala Phe Gln Pro Pro Glu Ile     210 215 220 Ala Asn Gly Leu Asp Thr Phe Ser Gly Phe Lys Val Asp Ile Trp Ser 225 230 235 240 Ala Gly Val Thr Leu Tyr Asn Ile Thr Thr Gly Leu Tyr Pro Phe Glu                 245 250 255 Gly Asp Asn Ile Tyr Lys Leu Phe Glu Asn Ile Gly Lys Gly Ser Tyr             260 265 270 Ala Ile Pro Gly Asp Cys Gly Pro Pro Leu Ser Asp Leu Leu Lys Gly         275 280 285 Met Leu Glu Tyr Glu Pro Ala Lys Arg Phe Ser Ile Arg Gln Ile Arg     290 295 300 Gln His Ser Trp Phe Arg Lys Lys His Pro Pro Ala Glu Ala Pro Val 305 310 315 320 Pro Ile Pro Pro Ser Pro Asp Thr Lys Asp Arg Trp Arg Ser Met Thr                 325 330 335 Val Val Pro Tyr Leu Glu Asp Leu His Gly Ala Asp Glu Asp Glu Asp             340 345 350 Leu Phe Asp Ile Glu Asp Asp Ile Ile Tyr Thr Gln Asp Phe Thr Val         355 360 365 Pro Gly Gln Val Pro Glu Glu Glu Ala Ser His Asn Gly Gln Arg Arg     370 375 380 Gly Leu Pro Lys Ala Val Cys Met Asn Gly Thr Glu Ala Ala Gln Leu 385 390 395 400 Ser Thr Lys Ser Arg Ala Glu Gly Arg Ala Pro Asn Pro Ala Arg Lys                 405 410 415 Ala Cys Ser Ala Ser Ser Lys Ile Arg Arg Leu Ser Ala Cys Lys Gln             420 425 430 Gln     <210> 27 <211> 156 <212> PRT <213> Homo sapiens_ CDKN2A amino acid <400> 27 Met Glu Pro Ala Ala Gly Ser Ser Met Glu Pro Ser Ala Asp Trp Leu   1 5 10 15 Ala Thr Ala Ala Ala Arg              20 25 30 Glu Ala Gly Ala Leu Pro Asn Ala Pro Asn Ser Tyr Gly Arg Arg Pro          35 40 45 Ile Gln Val Met Met Gly Ser Ala Arg Val Ala Glu Leu Leu Leu      50 55 60 Leu His Gly Ala Glu Pro Asn Cys Ala Asp Pro Ala Thr Leu Thr Arg  65 70 75 80 Pro Val His Asp Ala Ala Arg Glu Gly Phe Leu Asp Thr Leu Val Val                  85 90 95 Leu His Arg Ala Gly Ala Arg Leu Asp Val Arg Asp Ala Trp Gly Arg             100 105 110 Leu Pro Val Asp Leu Ala Glu Glu Leu Gly His Arg Asp Val Ala Arg         115 120 125 Tyr Leu Arg Ala Ala Ala Gly Gly Thr Arg Gly Ser Asn His Ala Arg     130 135 140 Ile Asp Ala Ala Glu Gly Pro Ser Asp Ile Pro Asp 145 150 155 <210> 28 <211> 552 <212> PRT <213> Homo sapiens SMAD4 amino acid <400> 28 Met Asp Asn Met Ser Ile Thr Asn Thr Pro Thr Ser Asn Asp Ala Cys   1 5 10 15 Leu Ser Ile Val His Ser Leu Met Cys His Arg Gln Gly Gly Glu Ser              20 25 30 Glu Thr Phe Ala Lys Arg Ala Ile Glu Ser Leu Val Lys Lys Leu Lys          35 40 45 Glu Lys Lys Asp Glu Leu Asp Ser Leu Ile Thr Ala Ile Thr Thr Asn      50 55 60 Gly Ala His Pro Ser Lys Cys Val Thr Ile Gln Arg Thr Leu Asp Gly  65 70 75 80 Arg Leu Gln Val Ala Gly Arg Lys Gly Phe Pro His Val Ile Tyr Ala                  85 90 95 Arg Leu Trp Arg Trp Pro Asp Leu His Lys Asn Glu Leu Lys His Val             100 105 110 Lys Tyr Cys Gln Tyr Ala Phe Asp Leu Lys Cys Asp Ser Val Cys Val         115 120 125 Asn Pro Tyr His Tyr Glu Arg Val Val Ser Ser Gly Ile Asp Leu Ser     130 135 140 Gly Leu Thr Leu Gln Ser Asn Ala Pro Ser Ser Met Met Val Lys Asp 145 150 155 160 Glu Tyr Val His Asp Phe Glu Gly Gln Pro Ser Leu Ser Thr Glu Gly                 165 170 175 His Ser Ile Gln Thr Ile Gln His Pro Ser Ser Asn Arg Ala Ser Thr             180 185 190 Glu Thr Tyr Ser Thr Pro Ala Leu Leu Ala Pro Ser Glu Ser Asn Ala         195 200 205 Thr Ser Thr Ala Asn Phe Pro Asn Ile Pro Val Ala Ser Thr Ser Gln     210 215 220 Pro Ala Ser Ile Leu Gly Gly Ser His Ser Glu Gly Leu Leu Gln Ile 225 230 235 240 Ala Ser Gly Pro Gln Pro Gly Gln Gln Gln Asn Gly Phe Thr Gly Gln                 245 250 255 Pro Ala Thr Tyr His His Ser Thr Thr Thr Trp Thr Gly Ser Arg             260 265 270 Thr Ala Pro Tyr Thr Pro Asn Leu Pro His His Gln Asn Gly His Leu         275 280 285 Gln His His Pro Met Pro Pro His Pro Gly His Tyr Trp Pro Val     290 295 300 His Asn Glu Leu Ala Phe Gln Pro Pro Ile Ser Asn His Pro Ala Pro 305 310 315 320 Glu Tyr Trp Cys Ser Ile Ala Tyr Phe Glu Met Asp Val Gln Val Gly                 325 330 335 Glu Thr Phe Lys Val Ser Ser Cys Pro Ile Val Thr Val Asp Gly             340 345 350 Tyr Val Asp Pro Ser Gly Gly Asp Arg Phe Cys Leu Gly Gln Leu Ser         355 360 365 Asn Val His Arg Thr Glu Ala Ile Glu Arg Ala Arg Leu His Ile Gly     370 375 380 Lys Gly Val Gln Leu Glu Cys Lys Gly Glu Gly Asp Val Trp Val Arg 385 390 395 400 Cys Leu Ser Asp His Ala Val Phe Val Gln Ser Tyr Tyr Leu Asp Arg                 405 410 415 Glu Ala Gly Arg Ala Pro Gly Asp Ala Val His Lys Ile Tyr Pro Ser             420 425 430 Ala Tyr Ile Lys Val Phe Asp Leu Arg Gln Cys His Arg Gln Met Gln         435 440 445 Gln Gln Ala Ala Thr Ala Gln Ala Ala Ala Ala Ala Gln Ala Ala Ala     450 455 460 Val Ala Gly Asn Ile Pro Gly Pro Gly Ser Val Gly Gly Ile Ala Pro 465 470 475 480 Ala Ile Ser Leu Ser Ala Ala Aly Gly Ile Aly Gly Val Asp Asp Leu Arg                 485 490 495 Arg Leu Cys Ile Leu Arg Met Ser Phe Val Lys Gly Trp Gly Pro Asp             500 505 510 Tyr Pro Arg Gln Ser Ile Lys Glu Thr Pro Cys Trp Ile Glu Ile His         515 520 525 Leu His Arg Ala Leu Gln Leu Leu Asp Glu Val Leu His Thr Met Pro     530 535 540 Ile Ala Asp Pro Gln Pro Leu Asp 545 550 <210> 29 <211> 2591 <212> DNA <213> Homo sapiens_TP53 gene <400> 29 gatgggattg gggttttccc ctcccatgtg ctcaagactg gcgctaaaag ttttgagctt 60 ctcaaaagtc tagagccacc gtccagggag caggtagctg ctgggctccg gggacacttt 120 gcgttcgggc tgggagcgtg ctttccacga cggtgacacg cttccctgga ttggcagcca 180 gactgccttc cgggtcactg ccatggagga gccgcagtca gatcctagcg tcgagccccc 240 tctgagtcag gaaacatttt cagacctatg gaaactactt cctgaaaaca acgttctgtc 300 ccccttgccg tccgaagcaa tggatgattt gatgctgtcc ccggacgata ttgaacaatg 360 gttcactgaa gacccaggtc cagatgaagc tcccagaatg ccagaggctg ctccccccgt 420 ggcccctgca ccagcagctc ctacaccggc ggcccctgca ccagccccct cctggcccct 480 gtcatcttct gtcccttccc agaaaaccta ccagggcagc tacggtttcc gtctgggctt 540 cttgcattct gggacagcca agtctgtgac ttgcacgtac tcccctgccc tcaacaagat 600 gtttggccaa ctggccaaga cctgccctgt gcagctgtgg gttgattcca cacccccgcc 660 cggcacccgc gtccgcgcca tggccatcta caagcagtca cagcacatga cggaggttgt 720 gaggcgctgc ccccaccatg agcgctgctc agatagcgat ggtctggccc ctcctcagca 780 tcttatccga gtggaaggaa atttgcgtgt ggagtatttg gatgacagaa acacttttcg 840 acatagtgtg gtggtgccct atgagccgcc tgaggttggc tctgactgta ccaccatcca 900 ctacaactac atgtgtaaca gttcctgcat gggcggcatg aaccggaggc ccatcctcac 960 catcatcaca ctggaagact ccagtggtaa tctactggga cggaacagct ttgaggtgcg 1020 tgtttgtgcc tgtcctggga gagaccggcg cacagaggaa gagaatctcc gcaagaaagg 1080 ggagcctcac cacgagctgc ccccagggag cactaagcga gcactgccca acaacaccag 1140 ctcctctccc cagccaaaga agaaaccact ggatggagaa tatttcaccc ttcagatccg 1200 tgggcgtgag cgcttcgaga tgttccgaga gctgaatgag gccttggaac tcaaggatgc 1260 ccaggctggg aaggagccag gggggagcag ggctcactcc agccacctga agtccaaaaa 1320 gggtcagtct acctcccgcc ataaaaaact catgttcaag acagaagggc ctgactcaga 1380 ctgacattct ccacttcttg ttccccactg acagcctccc acccccatct ctccctcccc 1440 tgccattttg ggttttgggt ctttgaaccc ttgcttgcaa taggtgtgcg tcagaagcac 1500 ccaggacttc catttgcttt gtcccggggc tccactgaac aagttggcct gcactggtgt 1560 tttgttgtgg ggaggaggat ggggagtagg acataccagc ttagatttta aggtttttac 1620 tgtgagggat gtttgggaga tgtaagaaat gttcttgcag ttaagggtta gtttacaatc 1680 agccacattc taggtagggg cccacttcac cgtactaacc agggaagctg tccctcactg 1740 ttgaattttc tctaacttca aggcccatat ctgtgaaatg ctggcatttg cacctacctc 1800 acagagtgca ttgtgagggt taatgaaata atgtacatct ggccttgaaa ccacctttta 1860 ttacatgggg tctagaactt gacccccttg agggtgcttg ttccctctcc ctgttggtcg 1920 gtgggttggt agtttctaca gttgggcagc tggttaggta gagggagttg tcaagtctct 1980 gctggcccag ccaaaccctg tctgacaacc tcttggtgaa ccttagtacc taaaaggaaa 2040 tctcacccca tcccacaccc tggaggattt catctcttgt atatgatgat ctggatccac 2100 caagacttgt tttatgctca gggtcaattt cttttttctt tttttttttt ttttttcttt 2160 ttctttgaga ctgggtctcg ctttgttgcc caggctggag tggagtggcg tgatcttggc 2220 ttactgcagc ctttgcctcc ccggctcgag cagtcctgcc tcagcctccg gagtagctgg 2280 gaccacaggt tcatgccacc atggccagcc aacttttgca tgttttgtag agatggggtc 2340 tcacagtgtt gcccaggctg gtctcaaact cctgggctca ggcgatccac ctgtctcagc 2400 ctcccagagt gctgggatta caattgtgag ccaccacgtc cagctggaag ggtcaacatc 2460 ttttacattc tgcaagcaca tctgcatttt caccccaccc ttcccctcct tctccctttt 2520 tatatcccat ttttatatcg atctcttatt ttacaataaa actttgctgc cacctgtgtg 2580 tctgaggggt g 2591 <210> 30 <211> 10740 <212> DNA <213> Homo sapiens_ APC gene <400> 30 gtattggtgc agcccgccag ggtgtcactg gagacagaat ggaggtgctg ccggactcgg 60 aaatggggtc caagggtagc caaggatggc tgcagcttca tatgatcagt tgttaaagca 120 agttgaggca ctgaagatgg agaactcaaa tcttcgacaa gagctagaag ataattccaa 180 tcatcttaca aaactggaaa ctgaggcatc taatatgaag gaagtactta aacaactaca 240 aggaagtatt gaagatgaag ctatggcttc ttctggacag attgatttat tagagcgtct 300 taaagagctt aacttagata gcagtaattt ccctggagta aaactgcggt caaaaatgtc 360 cctccgttct tatggaagcc gggaaggatc tgtatcaagc cgttctggag agtgcagtcc 420 tgttcctatg ggttcatttc caagaagagg gtttgtaaat ggaagcagag aaagtactgg 480 atatttagaa gaacttgaga aagagaggtc attgcttctt gctgatcttg acaaagaaga 540 aaaggaaaaa gactggtatt acgctcaact tcagaatctc actaaaagaa tagatagtct 600 tcctttaact gaaaattttt ccttacaaac agatatgacc agaaggcaat tggaatatga 660 agcaaggcaa atcagagttg cgatggaaga acaactaggt acctgccagg atatggaaaa 720 acgagcacag cgaagaatag ccagaattca gcaaatcgaa aaggacatac ttcgtatacg 780 acagctttta cagtcccaag caacagaagc agagaggtca tctcagaaca agcatgaaac 840 cggctcacat gatgctgagc ggcagaatga aggtcaagga gtgggagaaa tcaacatggc 900 aacttctggt aatggtcagg gttcaactac acgaatggac catgaaacag ccagtgtttt 960 gagttctagt agcacacact ctgcacctcg aaggctgaca agtcatctgg gaaccaaggt 1020 ggaaatggtg tattcattgt tgtcaatgct tggtactcat gataaggatg atatgtcgcg 1080 aactttgcta gctatgtcta gctcccaaga cagctgtata tccatgcgac agtctggatg 1140 tcttcctctc ctcatccagc ttttacatgg caatgacaaa gactctgtat tgttgggaaa 1200 ttcccggggc agtaaagagg ctcgggccag ggccagtgca gcactccaca acatcattca 1260 ctcacagcct gatgacaaga gaggcaggcg tgaaatccga gtccttcatc ttttggaaca 1320 gatacgcgct tactgtgaaa cctgttggga gtggcaggaa gctcatgaac caggcatgga 1380 ccaggacaaa aatccaatgc cagctcctgt tgaacatcag atctgtcctg ctgtgtgtgt 1440 tctaatgaaa ctttcatttg atgaagagca tagacatgca atgaatgaac tagggggact 1500 acaggccatt gcagaattat tgcaagtgga ctgtgaaatg tatgggctta ctaatgacca 1560 ctacagtatt acactaagac gatatgctgg aatggctttg acaaacttga cttttggaga 1620 tgtagccaac aaggctacgc tatgctctat gaaaggctgc atgagagcac ttgtggccca 1680 actaaaatct gaaagtgaag acttacagca ggttattgcg agtgttttga ggaatttgtc 1740 ttggcgagca gatgtaaata gtaaaaagac gttgcgagaa gttggaagtg tgaaagcatt 1800 gatggaatgt gctttagaag ttaaaaagga atcaaccctc aaaagcgtat tgagtgcctt 1860 atggaatttg tcagcacatt gcactgagaa taaagctgat atatgtgctg tagatggtgc 1920 acttgcattt ttggttggca ctcttactta ccggagccag acaaacactt tagccattat 1980 tgaaagtgga ggtgggatat tacggaatgt gtccagcttg atagctacaa atgaggacca 2040 caggcaaatc ctaagagaga acaactgtct acaaacttta ttacaacact taaaatctca 2100 tagtttgaca atagtcagta atgcatgtgg aactttgtgg aatctctcag caagaaatcc 2160 taaagaccag gaagcattat gggacatggg ggcagttagc atgctcaaga acctcattca 2220 ttcaaagcac aaaatgattg ctatgggaag tgctgcagct ttaaggaatc tcatggcaaa 2280 taggcctgcg aagtacaagg atgccaatat tatgtctcct ggctcaagct tgccatctct 2340 tcatgttagg aaacaaaaag ccctagaagc agaattagat gctcagcact tatcagaaac 2400 ttttgacaat atagacaatt taagtcccaa ggcatctcat cgtagtaagc agagacacaa 2460 gcaaagtctc tatggtgatt atgtttttga caccaatcga catgatgata ataggtcaga 2520 caattttaat actggcaaca tgactgtcct ttcaccatat ttgaatacta cagtgttacc 2580 cagctcctct tcatcaagag gaagcttaga tagttctcgt tctgaaaaag atagaagttt 2640 ggagagagaa cgcggaattg gtctaggcaa ctaccatcca gcaacagaaa atccaggaac 2700 ttcttcaaag cgaggtttgc agatctccac cactgcagcc cagattgcca aagtcatgga 2760 agaagtgtca gccattcata cctctcagga agacagaagt tctgggtcta ccactgaatt 2820 acattgtgtg acagatgaga gaaatgcact tagaagaagc tctgctgccc atacacattc 2880 aaacacttac aatttcacta agtcggaaaa ttcaaatagg acatgttcta tgccttatgc 2940 caaattagaa tacaagagat cttcaaatga tagtttaaat agtgtcagta gtagtgatgg 3000 ttatggtaaa agaggtcaaa tgaaaccctc gattgaatcc tattctgaag atgatgaaag 3060 taagttttgc agttatggtc aatacccagc cgacctagcc cataaaatac atagtgcaaa 3120 tcatatggat gataatgatg gagaactaga tacaccaata aattatagtc ttaaatattc 3180 agatgagcag ttgaactctg gaaggcaaag tccttcacag aatgaaagat gggcaagacc 3240 caaacacata atagaagatg aaataaaaca aagtgagcaa agacaatcaa ggaatcaaag 3300 tacaacttat cctgtttata ctgagagcac tgatgataaa cacctcaagt tccaaccaca 3360 ttttggacag caggaatgtg tttctccata caggtcacgg ggagccaatg gttcagaaac 3420 aaatcgagtg ggttctaatc atggaattaa tcaaaatgta agccagtctt tgtgtcaaga 3480 agatgactat gaagatgata agcctaccaa ttatagtgaa cgttactctg aagaagaaca 3540 gcatgaagaa gaagagagac caacaaatta tagcataaaa tataatgaag agaaacgtca 3600 tgtggatcag cctattgatt atagtttaaa atatgccaca gatattcctt catcacagaa 3660 acagtcattt tcattctcaa agagttcatc tggacaaagc agtaaaaccg aacatatgtc 3720 ttcaagcagt gagaatacgt ccacaccttc atctaatgcc aagaggcaga atcagctcca 3780 tccaagttct gcacagagta gaagtggtca gcctcaaaag gctgccactt gcaaagtttc 3840 ttctattaac caagaaacaa tacagactta ttgtgtagaa gatactccaa tatgtttttc 3900 aagatgtagt tcattatcat ctttgtcatc agctgaagat gaaataggat gtaatcagac 3960 gacacaggaa gcagattctg ctaataccct gcaaatagca gaaataaaag aaaagattgg 4020 aactaggtca gctgaagatc ctgtgagcga agttccagca gtgtcacagc accctagaac 4080 caaatccagc agactgcagg gttctagttt atcttcagaa tcagccaggc acaaagctgt 4140 tgaattttct tcaggagcga aatctccctc caaaagtggt gctcagacac ccaaaagtcc 4200 acctgaacac tatgttcagg agaccccact catgtttagc agatgtactt ctgtcagttc 4260 acttgatagt tttgagagtc gttcgattgc cagctccgtt cagagtgaac catgcagtgg 4320 aatggtaagt ggcattataa gccccagtga tcttccagat agccctggac aaaccatgcc 4380 accaagcaga agtaaaacac ctccaccacc tcctcaaaca gctcaaacca agcgagaagt 4440 acctaaaaat aaagcaccta ctgctgaaaa gagagagagt ggacctaagc aagctgcagt 4500 aaatgctgca gttcagaggg tccaggttct tccagatgct gatactttat tacattttgc 4560 cccggaaagt actccagatg gattttcttg ttcatccagc ctgagtgctc tgagcctcga 4620 tgagccattt atacagaaag atgtggaatt aagaataatg cctccagttc aggaaaatga 4680 caatgggaat gaaacagaat cagagcagcc taaagaatca aatgaaaacc aagagaaaga 4740 ggcagaaaaa actattgatt ctgaaaagga cctattagat gattcagatg atgatgatat 4800 tgaaatacta gaagaatgta ttatttctgc catgccaaca aagtcatcac gtaaagcaaa 4860 aaagccagcc cagactgctt caaaattacc tccacctgtg gcaaggaaac caagtcagct 4920 gcctgtgtac aaacttctac catcacaaaa caggttgcaa ccccaaaagc atgttagttt 4980 tacaccgggg gatgatatgc cacgggtgta ttgtgttgaa gggacaccta taaacttttc 5040 cacagctaca tctctaagtg atctaacaat cgaatcccct ccaaatgagt tagctgctgg 5100 agaaggagtt agaggagggg cacagtcagg tgaatttgaa aaacgagata ccattcctac 5160 agaaggcaga agtacagatg aggctcaagg aggaaaaacc tcatctgtaa ccatacctga 5220 attggatgac aataaagcag aggaaggtga tattcttgca gaatgcatta attctgctat 5280 gcccaaaggg aaaagtcaca agcctttccg tgtgaaaaag ataatggacc aggtccagca 5340 agcatctgcg tcttcttctg cacccaacaa aaatcagtta gatggtaaga aaaagaaacc 5400 aacttcacca gtaaaaccta taccacaaaa tactgaatat aggacacgtg taagaaaaaa 5460 tgcagactca aaaaataatt taaatgctga gagagttttc tcagacaaca aagattcaaa 5520 gaaacagaat ttgaaaaata attccaaggt cttcaatgat aagctcccaa ataatgaaga 5580 tagagtcaga ggaagttttg cttttgattc acctcatcat tacacgccta ttgaaggaac 5640 tccttactgt ttttcacgaa atgattcttt gagttctcta gattttgatg atgatgatgt 5700 tgacctttcc agggaaaagg ctgaattaag aaaggcaaaa gaaaataagg aatcagaggc 5760 taaagttacc agccacacag aactaacctc caaccaacaa tcagctaata agacacaagc 5820 tattgcaaag cagccaataa atcgaggtca gcctaaaccc atacttcaga aacaatccac 5880 ttttccccag tcatccaaag acataccaga cagaggggca gcaactgatg aaaagttaca 5940 gaattttgct attgaaaata ctccggtttg cttttctcat aattcctctc tgagttctct 6000 cagtgacatt gaccaagaaa acaacaataa agaaaatgaa cctatcaaag agactgagcc 6060 ccctgactca cagggagaac caagtaaacc tcaagcatca ggctatgctc ctaaatcatt 6120 tcatgttgaa gataccccag tttgtttctc aagaaacagt tctctcagtt ctcttagtat 6180 tgactctgaa gatgacctgt tgcaggaatg tataagctcc gcaatgccaa aaaagaaaaa 6240 gccttcaaga ctcaagggtg ataatgaaaa acatagtccc agaaatatgg gtggcatatt 6300 aggtgaagat ctgacacttg atttgaaaga tatacagaga ccagattcag aacatggtct 6360 atcccctgat tcagaaaatt ttgattggaa agctattcag gaaggtgcaa attccatagt 6420 aagtagttta catcaagctg ctgctgctgc atgtttatct agacaagctt cgtctgattc 6480 agattccatc ctttccctga aatcaggaat ctctctggga tcaccatttc atcttacacc 6540 tgatcaagaa gaaaaaccct ttacaagtaa taaaggccca cgaattctaa aaccagggga 6600 gaaaagtaca ttggaaacta aaaagataga atctgaaagt aaaggaatca aaggaggaaa 6660 aaaagtttat aaaagtttga ttactggaaa agttcgatct aattcagaaa tttcaggcca 6720 aatgaaacag ccccttcaag caaacatgcc ttcaatctct cgaggcagga caatgattca 6780 tattccagga gttcgaaata gctcctcaag tacaagtcct gtttctaaaa aaggcccacc 6840 ccttaagact ccagcctcca aaagccctag tgaaggtcaa acagccacca cttctcctag 6900 aggagccaag ccatctgtga aatcagaatt aagccctgtt gccaggcaga catcccaaat 6960 aggtgggtca agtaaagcac cttctagatc aggatctaga gattcgaccc cttcaagacc 7020 tgcccagcaa ccattaagta gacctataca gtctcctggc cgaaactcaa tttcccctgg 7080 tagaaatgga ataagtcctc ctaacaaatt atctcaactt ccaaggacat catcccctag 7140 tactgcttca actaagtcct caggttctgg aaaaatgtca tatacatctc caggtagaca 7200 gatgagccaa cagaacctta ccaaacaaac aggtttatcc aagaatgcca gtagtattcc 7260 aagaagtgag tctgcctcca aaggactaaa tcagatgaat aatggtaatg gagccaataa 7320 aaaggtagaa ctttctagaa tgtcttcaac taaatcaagt ggaagtgaat ctgatagatc 7380 agaaagacct gtattagtac gccagtcaac tttcatcaaa gaagctccaa gcccaacctt 7440 aagaagaaaa ttggaggaat ctgcttcatt tgaatctctt tctccatcat ctagaccagc 7500 ttctcccact aggtcccagg cacaaactcc agttttaagt ccttcccttc ctgatatgtc 7560 tctatccaca cattcgtctg ttcaggctgg tggatggcga aaactcccac ctaatctcag 7620 tcccactata gagtataatg atggaagacc agcaaagcgc catgatattg cacggtctca 7680 ttctgaaagt ccttctagac ttccaatcaa taggtcagga acctggaaac gtgagcacag 7740 caaacattca tcatcccttc ctcgagtaag cacttggaga agaactggaa gttcatcttc 7800 aattctttct gcttcatcag aatccagtga aaaagcaaaa agtgaggatg aaaaacatgt 7860 gaactctatt tcaggaacca aacaaagtaa agaaaaccaa gtatccgcaa aaggaacatg 7920 gagaaaaata aaagaaaatg aattttctcc cacaaatagt acttctcaga ccgtttcctc 7980 aggtgctaca aatggtgctg aatcaaagac tctaatttat caaatggcac ctgctgtttc 8040 taaaacagag gatgtttggg tgagaattga ggactgtccc attaacaatc ctagatctgg 8100 aagatctccc acaggtaata ctcccccggt gattgacagt gtttcagaaa aggcaaatcc 8160 aaacattaaa gattcaaaag ataatcaggc aaaacaaaat gtgggtaatg gcagtgttcc 8220 catgcgtacc gtgggtttgg aaaatcgcct gaactccttt attcaggtgg atgcccctga 8280 ccaaaaagga actgagataa aaccaggaca aaataatcct gtccctgtat cagagactaa 8340 tgaaagttct atagtggaac gtaccccatt cagttctagc agctcaagca aacacagttc 8400 acctagtggg actgttgctg ccagagtgac tccttttaat tacaacccaa gccctaggaa 8460 aagcagcgca gatagcactt cagctcggcc atctcagatc ccaactccag tgaataacaa 8520 cacaaagaag cgagattcca aaactgacag cacagaatcc agtggaaccc aaagtcctaa 8580 gcgccattct gggtcttacc ttgtgacatc tgtttaaaag agaggaagaa tgaaactaag 8640 aaaattctat gttaattaca actgctatat agacattttg tttcaaatga aactttaaaa 8700 gactgaaaaa ttttgtaaat aggtttgatt cttgttagag ggtttttgtt ctggaagcca 8760 tatttgatag tatactttgt cttcactggt cttattttgg gaggcactct tgatggttag 8820 gaaaaaaata gtaaagccaa gtatgtttgt acagtatgtt ttacatgtat ttaaagtagc 8880 atcccatccc aacttccttt aattattgct tgtcttaaaa taatgaacac tacagataga 8940 aaatatgata tattgctgtt atcaatcatt tctagattat aaactgacta aacttacatc 9000 agggaaaaat tggtatttat gcaaaaaaaa atgtttttgt ccttgtgagt ccatctaaca 9060 tcataattaa tcatgtggct gtgaaattca cagtaatatg gttcccgatg aacaagttta 9120 cccagcctgc tttgctttac tgcatgaatg aaactgatgg ttcaatttca gaagtaatga 9180 ttaacagtta tgtggtcaca tgatgtgcat agagatagct acagtgtaat aatttacact 9240 attttgtgct ccaaacaaaa caaaaatctg tgtaactgta aaacattgaa tgaaactatt 9300 ttacctgaac tagattttat ctgaaagtag gtagaatttt tgctatgctg taatttgttg 9360 tatattctgg tatttgaggt gagatggctg ctcttttatt aatgagacat gaattgtgtc 9420 tcaacagaaa ctaaatgaac atttcagaat aaattattgc tgtatgtaaa ctgttactga 9480 aattggtatt tgtttgaagg gtcttgtttc acatttgtat taataattgt ttaaaatgcc 9540 tcttttaaaa gcttatataa atttttttct tcagcttcta tgcattaaga gtaaaattcc 9600 tcttactgta ataaaaacaa ttgaagaaga ctgttgccac ttaaccattc catgcgttgg 9660 cacttatcta ttcctgaaat ttcttttatg tgattagctc atcttgattt ttaatatttt 9720 tccacttaaa cttttttttc ttactccact ggagctcagt aaaagtaaat tcatgtaata 9780 gcaatgcaag cagcctagca cagactaagc attgagcata ataggcccac ataatttcct 9840 ctttcttaat attatagaat tctgtacttg aaattgattc ttagacattg cagtctcttc 9900 gggctttac agtgtaaact gtcttgcccc ttcatcttct tgttgcaact gggtctgaca 9960 tgaacacttt ttatcaccct gtatgttagg gcaagatctc agcagtgaag tataatcagc 10020 actttgccat gctcagaaaa ttcaaatcac atggaacttt agaggtagat ttaatacgat 10080 taagatattc agaagtatat tttagaatcc ctgcctgtta aggaaacttt atttgtggta 10140 ggtacagttc tggggtacat gttaagtgtc cccttataca gtggagggaa gtcttccttc 10200 ctgaaggaaa ataaactgac acttattaac taagataatt tacttaatat atcttccctg 10260 atttgtttta aaagatcaga gggtgactga tgatacatgc atacatattt gttgaataaa 10320 tgaaaattta tttttagtga taagattcat acactctgta tttggggagg gaaaaccttt 10380 ttaagcatgg tggggcactc agataggagt gaatacacct acctggtgcc ttgaaaatca 10440 catcaagtag ttaattatct accccttacc tgtgtttata acttccaggt aatgagaatg 10500 atttttttta aagctaaaat gccagtaaat aaaagtgcta tgacttgagc taagatattt 10560 gactccaatg cctgtactgt gtctactgca ccactttgta aacacttcaa tttactatct 10620 ttgaaatgat tgacctttaa atttttgcca aatgttatct gaaattgtct atgaatacca 10680 tctacttctg ttgttttccc aggcttccat aaacaatgga gatacatgca aaaaaaaaaa 10740                                                                        10740 <210> 31 <211> 3424 <212> DNA <213> Homo sapiens_ PIK3CA gene <400> 31 aggatcagaa caatgcctcc aagaccatca tcaggtgaac tgtggggcat ccacttgatg 60 cccccaagaa tcctagtgga atgtttacta ccaaatggaa tgatagtgac tttagaatgc 120 ctccgtgagg ctacattagt aactataaag catgaactat ttaaagaagc aagaaaatac 180 cctctccatc aacttcttca agatgaatct tcttacattt tcgtaagtgt tacccaagaa 240 gcagaaaggg aagaattttt tgatgaaaca agacgacttt gtgatcttcg gctttttcaa 300 ccatttttaa aagtaattga accagtaggc aaccgtgaag aaaagatcct caatcgagaa 360 attggttttg ctatcggcat gccagtgtgc gaatttgata tggttaaaga tcctgaagta 420 caggacttcc gaagaaatat tcttaatgtt tgtaaagaag ctgtggatct tagggatctt 480 aattcacctc atagtagagc aatgtatgtc tatccgccac atgtagaatc ttcaccagag 540 ctgccaaagc acatatataa taaattggat agaggccaaa taatagtggt gatttgggta 600 atagtttctc caaataatga caagcagaag tatactctga aaatcaacca tgactgtgtg 660 ccagaacaag taattgctga agcaatcagg aaaaaaacta gaagtatgtt gctatcatct 720 gaacaattaa aactctgtgt tttagaatat cagggcaagt acattttaaa agtgtgtgga 780 tgtgatgaat acttcctaga aaaatatcct ctgagtcagt ataagtatat aagaagctgt 840 ataatgcttg ggaggatgcc caatttgaag atgatggcta aagaaagcct ttattctcaa 900 ctgccaatgg actgttttac aatgccatct tattccagac gcatttccac agctacacca 960 tatatgaatg gagaaacatc tacaaaatcc ctttgggtta taaatagagc actcagaata 1020 aaaattcttt gtgcaaccta cgtgaatcta aatattcgag acattgacaa gatttatgtt 1080 cgaacaggta tctaccatgg aggagaaccc ttatgtgaca atgtgaacac tcaaagagta 1140 ccttgttcca atcccaggtg gaatgaatgg ctgaattatg atatatacat tcctgatctt 1200 cctcgtgctg ctcgactttg cctttccatt tgctctgtta aaggccgaaa gggtgctaaa 1260 gaggaacact gtccattggc atggggaaat ataaacttgt ttgattacac agacactcta 1320 gtatctggaa aaatggcttt gaatctttgg ccagtacctc atggattaga agatttgctg 1380 aaccctattg gtgttactgg atcaaatcca aataaagaaa ctccatgctt agagttggag 1440 tttgactggt tcagcagtgt ggtaaagttc ccagatatgt cagtgattga agagcatgcc 1500 aattggtctg tatcccgaga agcaggattt agctattccc acgcaggact gagtaacaga 1560 ctagctagag acaatgaatt aagggaaaat gacaaagaac agctcaaagc aatttctaca 1620 cgagatcctc tctctgaaat cactgagcag gagaaagatt ttctatggag tcacagacac 1680 tattgtgtaa ctatccccga aattctaccc aaattgcttc tgtctgttaa atggaattct 1740 agagatgaag tagcccagat gtattgcttg gtaaaagatt ggcctccaat caaacctgaa 1800 caggctatgg aacttctgga ctgtaattac ccagatccta tggttcgagg ttttgctgtt 1860 cggtgcttgg aaaaatattt aacagatgac aaactttctc agtatttaat tcagctagta 1920 caggtcctaa aatatgaaca atatttggat aacttgcttg tgagattttt actgaagaaa 1980 gcattgacta atcaaaggat tgggcacttt ttcttttggc atttaaaatc tgagatgcac 2040 aataaaacag ttagccagag gtttggcctg cttttggagt cctattgtcg tgcatgtggg 2100 atgtatttga agcacctgaa taggcaagtc gaggcaatgg aaaagctcat taacttaact 2160 gacattctca aacaggagag gaaggatgaa acacaaaagg tacagatgaa gtttttagtt 2220 gagcaaatga ggcgaccaga tttcatggat gccctacagg gcttgctgtc tcctctaaac 2280 cctgctcatc aactaggaaa cctcaggctt aaagagtgtc gaattatgtc ttctgcaaaa 2340 aggccactgt ggttgaattg ggagaaccca gacatcatgt cagagttact gtttcagaac 2400 aatgagatca tctttaaaaa tggggatgat ttacggcaag atatgctaac acttcaaatt 2460 attcgtatta tggaaaatat ctggcaaaat caaggtcttg atcttcgaat gttaccttat 2520 ggttgtctgt caatcggtga ctgtgtggga cttattgagg tggtgcgaaa ttctcacact 2580 attatgcaaa ttcagtgcaa aggcggcttg aaaggtgcac tgcagttcaa cagccacaca 2640 ctacatcagt ggctcaaaga caagaacaaa ggagaaatat atgatgcagc cattgacctg 2700 tttacacgtt catgtgctgg atactgtgta gctaccttca ttttgggaat tggagatcgt 2760 cacaatagta acatcatggt gaaagacgat ggacaactgt ttcatataga ttttggacac 2820 tttttggatc acaagaagaa aaaatttggt tataaacgag aacgtgtgcc atttgttttg 2880 acacaggatt tcttaatagt gattagtaaa ggagcccaag aatgcacaaa gacaagagaa 2940 tttgagaggt ttcaggagat gtgttacaag gcttatctag ctattcgaca gcatgccaat 3000 ctcttcataa atcttttctc aatgatgctt ggctctggaa tgccagaact acaatctttt 3060 gatgacattg catacattcg aaagacccta gccttagata aaactgagca agaggctttg 3120 gagtatttca tgaaacaaat gaatgatgca catcatggtg gctggacaac aaaaatggat 3180 tggatcttcc acacaattaa acagcatgca ttgaactgaa agataactga gaaaatgaaa 3240 gctcactctg gattccacac tgcactgtta ataactctca gcaggcaaag accgattgca 3300 taggaattgc acaatccatg aacagcatta gatttacagc aagaacagaa ataaaatact 3360 atataattta aataatgtaa acgcaaacag ggtttgatag cacttaaact agttcatttc 3420 aaaa 3424 <210> 32 <211> 5765 <212> DNA <213> Homo sapiens_ KRAS gene <400> 32 tcctaggcgg cggccgcggc ggcggaggca gcagcggcgg cggcagtggc ggcggcgaag 60 gtggcggcgg ctcggccagt actcccggcc cccgccattt cggactggga gcgagcgcgg 120 cgcaggcact gaaggcggcg gcggggccag aggctcagcg gctcccaggt gcgggagaga 180 ggcctgctga aaatgactga atataaactt gtggtagttg gagctggtgg cgtaggcaag 240 agtgccttga cgatacagct aattcagaat cattttgtgg acgaatatga tccaacaata 300 gaggattcct acaggaagca agtagtaatt gatggagaaa cctgtctctt ggatattctc 360 gacacagcag gtcaagagga gtacagtgca atgagggacc agtacatgag gactggggag 420 ggctttcttt gtgtatttgc cataaataat actaaatcat ttgaagatat tcaccattat 480 agagaacaaa ttaaaagagt taaggactct gaagatgtac ctatggtcct agtaggaaat 540 aaatgtgatt tgccttctag aacagtagac acaaaacagg ctcaggactt agcaagaagt 600 tatggaattc cttttattga aacatcagca aagacaagac agggtgttga tgatgccttc 660 tatacattag ttcgagaaat tcgaaaacat aaagaaaaga tgagcaaaga tggtaaaaag 720 aagaaaaaga agtcaaagac aaagtgtgta attatgtaaa tacaatttgt acttttttct 780 taaggcatac tagtacaagt ggtaattttt gtacattaca ctaaattatt agcatttgtt 840 ttagcattac ctaatttttt tcctgctcca tgcagactgt tagcttttac cttaaatgct 900 tattttaaaa tgacagtgga agtttttttt tcctctaagt gccagtattc ccagagtttt 960 ggtttttgaa ctagcaatgc ctgtgaaaaa gaaactgaat acctaagatt tctgtcttgg 1020 ggtttttggt gcatgcagtt gattacttct tatttttctt accaattgtg aatgttggtg 1080 tgaaacaaat taatgaagct tttgaatcat ccctattctg tgttttatct agtcacataa 1140 atggattaat tactaatttc agttgagacc ttctaattgg tttttactga aacattgagg 1200 gaacacaaat ttatgggctt cctgatgatg attcttctag gcatcatgtc ctatagtttg 1260 tcatccctga tgaatgtaaa gttacactgt tcacaaaggt tttgtctcct ttccactgct 1320 attagtcatg gtcactctcc ccaaaatatt atattttttc tataaaaaga aaaaaatgga 1380 aaaaaattac aaggcaatgg aaactattat aaggccattt ccttttcaca ttagataaat 1440 tactataaag actcctaata gcttttcctg ttaaggcaga cccagtatga aatggggatt 1500 attatagcaa ccattttggg gctatattta catgctacta aatttttata ataattgaaa 1560 agattttaac aagtataaaa aattctcata ggaattaaat gtagtctccc tgtgtcagac 1620 tgctctttca tagtataact ttaaatcttt tcttcaactt gagtctttga agatagtttt 1680 aattctgctt gtgacattaa aagattattt gggccagtta tagcttatta ggtgttgaag 1740 agaccaaggt tgcaaggcca ggccctgtgt gaacctttga gctttcatag agagtttcac 1800 agcatggact gtgtccccac ggtcatccag tgttgtcatg cattggttag tcaaaatggg 1860 gagggactag ggcagtttgg atagctcaac aagatacaat ctcactctgt ggtggtcctg 1920 ctgacaaatc aagagcattg cttttgtttc ttaagaaaac aaactctttt ttaaaaatta 1980 cttttaaata ttaactcaaa agttgagatt ttggggtggt ggtgtgccaa gacattaatt 2040 ttttttttaa acaatgaagt gaaaaagttt tacaatctct aggtttggct agttctctta 2100 acactggtta aattaacatt gcataaacac ttttcaagtc tgatccatat ttaataatgc 2160 tttaaaataa aaataaaaac aatccttttg ataaatttaa aatgttactt attttaaaat 2220 aaatgaagtg agatggcatg gtgaggtgaa agtatcactg gactaggaag aaggtgactt 2280 aggttctaga taggtgtctt ttaggactct gattttgagg acatcactta ctatccattt 2340 cttcatgtta aaagaagtca tctcaaactc ttagtttttt ttttttacaa ctatgtaatt 2400 tatattccat ttacataagg atacacttat ttgtcaagct cagcacaatc tgtaaatttt 2460 taacctatgt tacaccatct tcagtgccag tcttgggcaa aattgtgcaa gaggtgaagt 2520 ttatatttga atatccattc tcgttttagg actcttcttc catattagtg tcatcttgcc 2580 tccctacctt ccacatgccc catgacttga tgcagtttta atacttgtaa ttcccctaac 2640 cataagattt actgctgctg tggatatctc catgaagttt tcccactgag tcacatcaga 2700 aatgccctac atcttatttc ctcagggctc aagagaatct gacagatacc ataaagggat 2760 ttgacctaat cactaatttt caggtggtgg ctgatgcttt gaacatctct ttgctgccca 2820 atccattagc gacagtagga tttttcaaac ctggtatgaa tagacagaac cctatccagt 2880 ggaaggagaa tttaataaag atagtgctga aagaattcct taggtaatct ataactagga 2940 ctactcctgg taacagtaat acattccatt gttttagtaa ccagaaatct tcatgcaatg 3000 aaaaatactt taattcatga agcttacttt ttttttttgg tgtcagagtc tcgctcttgt 3060 cacccaggct ggaatgcagt ggcgccatct cagctcactg caacctccat ctcccaggtt 3120 caagcgattc tcgtgcctcg gcctcctgag tagctgggat tacaggcgtg tgccactaca 3180 ctcaactaat ttttgtattt ttaggagaga cggggtttca ccctgttggc caggctggtc 3240 tcgaactcct gacctcaagt gattcaccca ccttggcctc ataaacctgt tttgcagaac 3300 tcatttattc agcaaatatt tattgagtgc ctaccagatg ccagtcaccg cacaaggcac 3360 tgggtatatg gtatccccaa acaagagaca taatcccggt ccttaggtag tgctagtgtg 3420 gtctgtaata tcttactaag gcctttggta tacgacccag agataacacg atgcgtattt 3480 tagttttgca aagaaggggt ttggtctctg tgccagctct ataattgttt tgctacgatt 3540 ccactgaaac tcttcgatca agctacttta tgtaaatcac ttcattgttt taaaggaata 3600 aacttgatta tattgttttt ttatttggca taactgtgat ttttttagga caattactgt 3660 acacattaag gtgtatgtca gatattcata ttgacccaaa tgtgtaatat tccagttttc 3720 tctgcataag taattaaaat atacttaaaa attaatagtt ttatctgggt acaaataaac 3780 aggtgcctga actagttcac agacaaggaa acttctatgt aaaaatcact atgatttctg 3840 aattgctatg tgaaactaca gatctttgga acactgttta ggtagggtgt taagacttac 3900 acagtacctc gtttctacac agagaaagaa atggccatac ttcaggaact gcagtgctta 3960 tgaggggata tttaggcctc ttgaattttt gatgtagatg ggcatttttt taaggtagtg 4020 gttaattacc tttatgtgaa ctttgaatgg tttaacaaaa gatttgtttt tgtagagatt 4080 ttaaaggggg agaattctag aaataaatgt tacctaatta ttacagcctt aaagacaaaa 4140 atccttgttg aagttttttt aaaaaaagct aaattacata gacttaggca ttaacatgtt 4200 tgtggaagaa tatagcagac gtatattgta tcatttgagt gaatgttccc aagtaggcat 4260 tctaggctct atttaactga gtcacactgc ataggaattt agaacctaac ttttataggt 4320 tatcaaaact gttgtcacca ttgcacaatt ttgtcctaat atatacatag aaactttgtg 4380 gggcatgtta agttacagtt tgcacaagtt catctcattt gtattccatt gatttttttt 4440 ttcttctaaa cattttttct tcaaacagta tataactttt tttaggggat ttttttttag 4500 acagcaaaaa ctatctgaag atttccattt gtcaaaaagt aatgatttct tgataattgt 4560 gtagtaatgt tttttagaac ccagcagtta ccttaaagct gaatttatat ttagtaactt 4620 ctgtgttaat actggatagc atgaattctg cattgagaaa ctgaatagct gtcataaaat 4680 gaaactttct ttctaaagaa agatactcac atgagttctt gaagaatagt cataactaga 4740 ttaagatctg tgttttagtt taatagtttg aagtgcctgt ttgggataat gataggtaat 4800 ttagatgaat ttaggggaaa aaaaagttat ctgcagatat gttgagggcc catctctccc 4860 cccacacccc cacagagcta actgggttac agtgttttat ccgaaagttt ccaattccac 4920 tgtcttgtgt tttcatgttg aaaatacttt tgcatttttc ctttgagtgc caatttctta 4980 ctagtactat ttcttaatgt aacatgttta cctggaatgt attttaacta tttttgtata 5040 gtgtaaactg aaacatgcac attttgtaca ttgtgctttc ttttgtggga catatgcagt 5100 gtgatccagt tgttttccat catttggttg cgctgaccta ggaatgttgg tcatatcaaa 5160 cattaaaaat gaccactctt ttaattgaaa ttaactttta aatgtttata ggagtatgtg 5220 ctgtgaagtg atctaaaatt tgtaatattt ttgtcatgaa ctgtactact cctaattatt 5280 gtaatgtaat aaaaatagtt acagtgacta tgagtgtgta tttattcatg aaatttgaac 5340 tgtttgcccc gaaatggata tggaatactt tataagccat agacactata gtataccagt 5400 gt; tgtaaaggcg tgtttgctta aacttaaaac catatttaga agtagatgca aaacaaatct 5520 gcctttatga caaaaaaata ggataacatt atttatttat ttccttttat caaagaaggt 5580 aattgataca caacaggtga cttggtttta ggcccaaagg tagcagcagc aacattaata 5640 atggaaataa ttgaatagtt agttatgtat gttaatgcca gtcaccagca ggctatttca 5700 aggtcagaag taatgactcc atacatatta tttatttcta taactacatt taaatcatta 5760 ccagg 5765 <210> 33 <211> 3772 <212> DNA <213> Homo sapiens SMO gene <400> 33 gggctgaaga caacttggat tgcgaggcta gggcttgggg agtcgtgcat cccgttccgg 60 gcctccgcag cccaacatgg gccccgggtt ccaaagtttg cgaagttggg cgccgagggg 120 ccggggcgcg cggagcgtcc gggggggccc gggcccggat tctctgggcg cacaggtcgc 180 ctgagccgcc tccgcggccg ccgaggtcgt gcgtgtggcc ggggggctcc gaggagcagg 240 cgggggcgcc ggggcttttg ctgagttggc ggggttggcc atggccgctg cccgcccagc 300 gcgggggccg gagctcccgc tcctggggct gctgctgctg ctgctgctgg gggacccggg 360 ccggggggcg gcctcgagcg ggaacgcgac cgggcctggg cctcggagcg cgggcgggag 420 cgcgaggagg agcgcggcgg tgactggccc tccgccgccg ctgagccact gcggccgggc 480 tgccccctgc gagccgctgc gctacaacgt gtgcctgggc tcggtgctgc cctacggggc 540 cacctccaca ctgctggccg gagactcgga ctcccaggag gaagcgcacg gcaagctcgt 600 gctctggtcg ggcctccgga atgccccccg ctgctgggca gtgatccagc ccctgctgtg 660 tgccgtatac atgcccaagt gtgagaatga ccgggtggag ctgcccagcc gtaccctctg 720 ccaggccacc cgaggcccct gtgccatcgt ggagagggag cggggctggc ctgacttcct 780 gcgctgcact cctgaccgct tccctgaagg ctgcacgaat gaggtgcaga acatcaagtt 840 caacagttca ggccagtgcg aagtgccctt ggttcggaca gacaacccca agagctggta 900 cgaggacgtg gagggctgcg gcatccagtg ccagaacccg ctcttcacag aggctgagca 960 ccaggacatg cacagctaca tcgcggcctt cggggccgtc acgggcctct gcacgctctt 1020 caccctggcc acattcgtgg ctgactggcg gaactcgaat cgctaccctg ctgttattct 1080 cttctacgtc aatgcgtgct tctttgtggg cagcattggc tggctggccc agttcatgga 1140 tggtgcccgc cgagagatcg tctgccgtgc agatggcacc atgaggcttg gggagcccac 1200 ctccaatgag actctgtcct gcgtcatcat ctttgtcatc gtgtactacg ccctgatggc 1260 tggtgtggtt tggtttgtgg tcctcaccta tgcctggcac acttccttca aagccctggg 1320 caccacctac cagcctctct cgggcaagac ctcctacttc cacctgctca cctggtcact 1380 cccctttgtc ctcactgtgg caatccttgc tgtggcgcag gtggatgggg actctgtgag 1440 tggcatttgt tttgtgggct acaagaacta ccgataccgt gcgggcttcg tgctggcccc 1500 aatcggcctg gtgctcatcg tgggaggcta cttcctcatc cgaggagtca tgactctgtt 1560 ctccatcaag agcaaccacc ccgggctgct gagtgagaag gctgccagca agatcaacga 1620 gaccatgctg cgcctgggca tttttggctt cctggccttt ggctttgtgc tcattacctt 1680 cagctgccac ttctacgact tcttcaacca ggctgagtgg gagcgcagct tccgggacta 1740 tgtgctatgt caggccaatg tgaccatcgg gctgcccacc aagcagccca tccctgactg 1800 tgagatcaag aatcgcccga gccttctggt ggagaagatc aacctgtttg ccatgtttgg 1860 aactggcatc gccatgagca cctgggtctg gaccaaggcc acgctgctca tctggaggcg 1920 tacctggtgc aggttgactg ggcagagtga cgatgagcca aagcggatca agaagagcaa 1980 gatgattgcc aaggccttct ctaagcggca cgagctcctg cagaacccag gccaggagct 2040 gtccttcagc atgcacactg tgtcccacga cgggcccgtg gcgggcttgg cctttgacct 2100 caatgagccc tcagctgatg tctcctctgc ctgggcccag catgtcacca agatggtggc 2160 tcggagagga gccatactgc cccaggatat ttctgtcacc cctgtggcaa ctccagtgcc 2220 cccagaggaa caagccaacc tgtggctggt tgaggagag atctccccag agctgcagaa 2280 gcgcctgggc cggaagaaga agaggaggaa gaggaagaag gaggtgtgcc cgctggcgcc 2340 gccccctgag cttcaccccc ctgcccctgc ccccagtacc attcctcgac tgcctcagct 2400 gccccggcag aaatgcctgg tggctgcagg tgcctgggga gctggggact cttgccgaca 2460 gggagcgtgg accctggtct ccaacccatt ctgcccagag cccagtcccc ctcaggatcc 2520 atttctgccc agtgcaccgg cccccgtggc atgggctcat ggccgccgac agggcctggg 2580 gcctattcac tcccgcacca acctgatgga cacagaactc atggatgcag actcggactt 2640 ctgagcctgc agagcaggac ctgggacagg aaagagagga accaatacct tcaaggctct 2700 tcttcctcac cgagcatgct tccctaggat cccgtcttcc agagaacctg tgggctgact 2760 gccctccgaa gagagttctg gatgtctggc tcaaagcagc aggactgtgg gaaagagcct 2820 aacatctcca tggggaggcc tcaccccagg gacagggccc tggagctcag ggtccttgtt 2880 tctgccctgc cagctgcagc ctggttggca gcatctgctc catcggggca gggggtatgc 2940 agagcttgtg gtggggcagg aacggtggag gcagaggtga cagttcccag agtgggcttt 3000 ggtggccagg gaggcagcct agcctatgtc tggcagatga gggctggctg ccgttttctg 3060 ggctgatggg tgccctttcc tggcagtctc agtccaaaag tgttgactgt gtcattagtc 3120 ctttgtctaa gtagggccag ggcaccgtat tcctctccca ggtgtttgtg gggctggaag 3180 gacctgctcc cacaggggcc atgtcctctc ttaataggtg gcactacccc aaacccatct 3240 tttgttctcc tatatcctcc ttctcctgtt ccatttcagt tcagtttcag cggtgccaac 3300 ctctttgcgt ttcctttttg ttgatgagga cccagagctg ctgcacacac tcacctctaa 3360 ccccctcccc tcgctgctgg gccccatctc cacaggagag actggttcgg ctctagggcc 3420 tcagtctgga gtgggatagg agcagtgagt gacaaagcct ctgaaagatg catcatctct 3480 tcctcacacc catttagtgg gggatgggtc ctctagactt gaggggctac cctgggaagc 3540 tgccgtagct tcagccaggc aagaaagctt ccttcaacct gcatagccgg tgggtgagga 3600 gattcccacc ttccatagcc tccaaacatg ttcccaaggc cccactttca agaatcagac 3660 agcaggaagc catagatgct ggctgggttc caggttatgg ggagaagaaa tacagtcaat 3720 aaaaggtttt tgtataaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaaaa 3772 <210> 34 <211> 3286 <212> DNA <213> Homo sapiens STK11 gene <400> 34 gcgtgtcggg cgcggaaggg ggaggcggcc cggggcgccc gcgagtgagg cgcggggcgg 60 cgaagggagc gcgggtggcg gcacttgctg ccgcggcctt ggatgggctg ggcccccctc 120 gccgctccgc ctcctccaca cgcgcggcgg ccgcggcgag ggggacgcgc cgcccggggc 180 ccggcacctt cgggaacccc ccggcccgga gcctgcggcc tgcgccgcct cggccgccgg 240 gagccccgtg gagcccccgc cgccgcgccg ccccgcggac cggacgctga gggcactcgg 300 ggcggggcgc gcgctcgggc agacgtttgc ggggaggggg gcgcctgccg ggccccggcg 360 cgcgggcgcc 420 gggcagcgac cagccctgag cggagctgtt ggccgcggcg ggaggcctcc cggacgcccc 480 cagccccccg aacgctcgcc cgggccggcg ggagtcggcg ccccccggga ggtccgctcg 540 gtcgtccgcg gcggagcgtt tgctcctggg acaggcggtg ggaccggggc gtcgccggag 600 acgcccccag cgaagttggg ctctccaggt gtgggggtcc cggggggtag cgacgtcgcg 660 gacccggcct gtgggatggg cggcccggag aagactgcgc tcggccgtgt tcatacttgt 720 ccgtgggcct gaggtccccg gaggatgacc tagcactgaa aagccccggc cggcctcccc 780 agggtccccg aggacgaagt tgaccctgac cgggccgtct cccagttctg aggcccgggt 840 cccactggaa ctcgcgtctg agccgccgtc ccggaccccc ggtgcccgcc ggtccgcaga 900 ccctgcaccg ggcttggact cgcagccggg actgacgtgt agaacaatcg tttctgttgg 960 aagaagggtt tttcccttcc ttttggggtt tttgttgcct tttttttttc ttttttcttt 1020 gtaaaatttt ggagaaggga agtcggaaca caaggaagga ccgctcaccc gcggactcag 1080 ggctggcggc gggactccag gaccctgggt ccagcatgga ggtggtggac ccgcagcagc 1140 tgggcatgtt cacggagggc gagctgatgt cggtgggtat ggacacgttc atccaccgca 1200 tcgactccac cgaggtcatc taccagccgc gccgcaagcg ggccaagctc atcggcaagt 1260 acctgatggg ggacctgctg ggggaaggct cttacggcaa ggtgaaggag gtgctggact 1320 cggagacgct gtgcaggagg gccgtcaaga tcctcaagaa gaagaagttg cgaaggatcc 1380 ccaacgggga ggccaacgtg aagaaggaaa ttcaactact gaggaggtta cggcacaaaa 1440 atgtcatcca gctggtggat gtgttataca acgaagagaa gcagaaaatg tatatggtga 1500 tggagtactg cgtgtgtggc atgcaggaaa tgctggacag cgtgccggag aagcgtttcc 1560 cagtgtgcca ggcccacggg tacttctgtc agctgattga cggcctggag tacctgcata 1620 gccagggcat tgtgcacaag gacatcaagc cggggaacct gctgctcacc accggtggca 1680 ccctcaaaat ctccgacctg ggcgtggccg aggcactgca cccgttcgcg gcggacgaca 1740 cctgccggac cagccagggc tccccggctt tccagccgcc cgagattgcc aacggcctgg 1800 acaccttctc cggcttcaag gtggacatct ggtcggctgg ggtcaccctc tacaacatca 1860 ccacgggtct gtaccccttc gaaggggaca acatctacaa gttgtttgag aacatcggga 1920 aggggagcta cgccatcccg ggcgactgtg gccccccgct ctctgacctg ctgaaaggga 1980 tgcttgagta cgaaccggcc aagaggttct ccatccggca gatccggcag cacagctggt 2040 tccggaagaa acatcctccg gctgaagcac cagtgcccat cccaccgagc ccagacacca 2100 aggaccggtg gcgcagcatg actgtggtgc cgtacttgga ggacctgcac ggcgcggacg 2160 aggacgagga cctcttcgac atcgaggatg acatcatcta cactcaggac ttcacggtgc 2220 ccggacaggt cccagaagag gaggccagtc acaatggaca gcgccggggc ctccccaagg 2280 ccgtgtgtat gaacggcaca gaggcggcgc agctgagcac caaatccagg gcggagggcc 2340 gggcccccaa ccctgcccgc aaggcctgct ccgccagcag caagatccgc cggctgtcgg 2400 cctgcaagca gcagtgaggc tggccgcctg cagcccgtgt ccaggagccc cgccaggtgc 2460 ccgcgccagg ccctcagtct tcctgccggt tccgcccgcc ctcccggaga ggtggccgcc 2520 atgcttctgt gccgaccacg ccccaggacc tccggagcgc cctgcagggc cgggcagggg 2580 gacagcaggg accgggcgca gccctccccc ctcggccgcc cggcagtgca cgcggcttgt 2640 tgacttcgca gccccgggcg gagccttccc gggcgggcgt gggaggaggg aggcggcctc 2700 catgcacttt atgtggagac tactggcccc gcccgtggcc tcgtgctccg cagggcgccc 2760 agcgccgtcc ggcggccccg ccgcagacca gctggcgggt gtggagacca ggctcctgac 2820 cccgccatgc atgcagcgcc acctggaagc cgcgcggccg ctttggtttt ttgtttggtt 2880 ggttccattt tctttttttc tttttttttt taagaaaaaa taaaaggtgg atttgagctg 2940 tggctgtgag gggtgtttgg gagctgctgg gtggcagggg ggctgtgggg tcgggctcac 3000 gtcgcggccg cctttgcgct ctcgggtcac cctgctttgg cggcccggcc ggagggcagg 3060 accctcacct ctcccccaag gccactgcgc tcttgggacc ccagagaaaa cccggagcaa 3120 gcaggagtgt gcggtcaata tttatatcat ccagaaaaga aaaacacgag aaacgccatc 3180 gcgggatggt gcagacgcgg cggggactcg gagggtgccg tgcgggcgag gccgcccaaa 3240 tttggcaata aataaagctt gggaagcttg gacctgaaaa aaaaaa 3286 <210> 35 <211> 1267 <212> DNA <213> Homo sapiens_ CDKN2A gene <400> 35 cgagggctgc ttccggctgg tgcccccggg ggagacccaa cctggggcga cttcaggggt 60 gccacattcg ctaagtgctc ggagttaata gcacctcctc cgagcactcg ctcacggcgt 120 ccccttgcct ggaaagatac cgcggtccct ccagaggatt tgagggacag ggtcggaggg 180 ggctcttccg ccagcaccgg aggaagaaag aggaggggct ggctggtcac cagagggtgg 240 ggcggaccgc gtgcgctcgg cggctgcgga gagggggaga gcaggcagcg ggcggcgggg 300 agcagcatgg agccggcggc ggggagcagc atggagcctt cggctgactg gctggccacg 360 gccgcggccc ggggtcgggt agaggaggtg cgggcgctgc tggaggcggg ggcgctgccc 420 aacgcaccga atagttacgg tcggaggccg atccaggtca tgatgatggg cagcgcccga 480 gtggcggagc tgctgctgct ccacggcgcg gagcccaact gcgccgaccc cgccactctc 540 acccgacccg tgcacgacgc tgcccgggag ggcttcctgg acacgctggt ggtgctgcac 600 cgggccgggg cgcggctgga cgtgcgcgat gcctggggcc gtctgcccgt ggacctggct 660 gaggagctgg gccatcgcga tgtcgcacgg tacctgcgcg cggctgcggg gggcaccaga 720 ggcagtaacc atgcccgcat agatgccgcg gaaggtccct cagacatccc cgattgaaag 780 aaccagagag gctctgagaa acctcgggaa acttagatca tcagtcaccg aaggtcctac 840 agggccacaa ctgcccccgc cacaacccac cccgctttcg tagttttcat ttagaaaata 900 gagcttttaa aaatgtcctg ccttttaacg tagatatatg ccttccccca ctaccgtaaa 960 tgtccattta tatcattttt tatatattct tataaaaatg taaaaaagaa aaacaccgct 1020 tctgcctttt cactgtgttg gagttttctg gagtgagcac tcacgcccta agcgcacatt 1080 catgtgggca tttcttgcga gcctcgcagc ctccggaagc tgtcgacttc atgacaagca 1140 ttttgtgaac tagggaagct caggggggtt actggcttct cttgagtcac actgctagca 1200 aatggcagaa ccaaagctca aataaaaata aaataatttt cattcattca ctcaaaaaaa 1260 aaaaaaa 1267 <210> 36 <211> 3220 <212> DNA <213> Homo sapiens SMAD4 gene <400> 36 caacaacacg gccctggtcg tcgtcgccgc tgcggtaacg gagcggtttg ggtggcggag 60 cctgcgttcg cgccttccca ctcccctcgc caccgcccga gcccaggtta tcctgaatac 120 atgtctaaca attttccttg caacgttagc tgttgttttt cactgtttcc aaaggatcaa 180 aattgcttca gaaattggag acatatttga tttaaaagga aaaacttgaa caaatggaca 240 atatgtctat tacgaataca ccaacaagta atgatgcctg tctgagcatt gtgcatagtt 300 tgatgtgcca tagacaaggt ggagagagtg aaacatttgc aaaaagagca attgaaagtt 360 tggtaaagaa gctgaaggag aaaaaagatg aattggattc tttaataaca gctataacta 420 caaatggagc tcatcctagt aaatgtgtta ccatacagag aacattggat gggaggcttc 480 aggtggctgg tcggaaagga tttcctcatg tgatctatgc ccgtctctgg aggtggcctg 540 atcttcacaa aaatgaacta aaacatgtta aatattgtca gtatgcgttt gacttaaaat 600 gtgatagtgt ctgtgtgaat ccatatcact acgaacgagt tgtatcacct ggaattgatc 660 tctcaggatt aacactgcag agtaatgctc catcaagtat gatggtgaag gatgaatatg 720 tgcatgactt tgagggacag ccatcgttgt ccactgaagg acattcaatt caaaccatcc 780 agcatccacc aagtaatcgt gcatcgacag agacatacag caccccagct ctgttagccc 840 catctgagtc taatgctacc agcactgcca actttcccaa cattcctgtg gcttccacaa 900 gtcagcctgc cagtatactg gggggcagcc atagtgaagg actgttgcag atagcatcag 960 ggcctcagcc aggacagcag cagaatggat ttactggtca gccagctact taccatcata 1020 acagcactac cacctggact ggaagtagga ctgcaccata cacacctaat ttgcctcacc 1080 accaaaacgg ccatcttcag caccacccgc ctatgccgcc ccatcccgga cattactggc 1140 ctgttcacaa tgagcttgca ttccagcctc ccatttccaa tcatcctgct cctgagtatt 1200 ggtgttccat tgcttacttt gaaatggatg ttcaggtagg agagacattt aaggttcctt 1260 caagctgccc tattgttact gttgatggat acgtggaccc ttctggagga gatcgctttt 1320 gtttgggtca actctccaat gtccacagga cagaagccat tgagagagca aggttgcaca 1380 taggcaaagg tgtgcagttg gaatgtaaag gtgaaggtga tgtttgggtc aggtgcctta 1440 gtgaccacgc ggtctttgta cagagttact acttagacag agaagctggg cgtgcacctg 1500 gagatgctgt tcataagatc tacccaagtg catatataaa ggtctttgat ttgcgtcagt 1560 gtcatcgaca gatgcagcag caggcggcta ctgcacaagc tgcagcagct gcccaggcag 1620 cagccgtggc aggaaacatc cctggcccag gatcagtagg tggaatagct ccagctatca 1680 gtctgtcagc tgctgctgga attggtgttg atgaccttcg tcgcttatgc atactcagga 1740 tgagttttgt gaaaggctgg ggaccggatt acccaagaca gagcatcaaa gaaacacctt 1800 gctggattga aattcactta caccgggccc tccagctcct agacgaagta cttcatacca 1860 tgccgattgc agacccacaa cctttagact gaggtctttt accgttgggg cccttaacct 1920 tatcaggatg gtggactaca aaatacaatc ctgtttataa tctgaagata tatttcactt 1980 ttgttctgct ttatcttttc ataaagggtt gaaaatgtgt ttgctgcctt gctcctagca 2040 gacagaaact ggattaaaac aatttttttt ttcctcttca gaacttgtca ggcatggctc 2100 agagcttgaa gattaggaga aacacattct tattaattct tcacctgtta tgtatgaagg 2160 aatcattcca gtgctagaaa atttagccct ttaaaacgtc ttagagcctt ttatctgcag 2220 aacatcgata tgtatatcat tctacagaat aatccagtat tgctgatttt aaaggagag 2280 aagttctcaa agttaattca cctatgttat tttgtgtaca agttgttatt gttgaacata 2340 cttcaaaaat aatgtgccat gtgggtgagt taattttacc aagagtaact ttactctgtg 2400 tttaaaaagt aagttaataa tgtattgtaa tctttcatcc aaaatatttt ttgcaagtta 2460 tattagtgaa gatggtttca attcagattg tcttgcaact tcagttttat ttttgccaag 2520 gcaaaaaact cttaatctgt gtgtatattg agaatccctt aaaattacca gacaaaaaaa 2580 tttaaaatta cgtttgttat tcctagtgga tgactgttga tgaagtatac ttttcccctg 2640 ttaaacagta gttgtattct tctgtatttc taggcacaag gttggttgct aagaagccta 2700 taagaggaat ttcttttcct tcattcatag ggaaaggttt tgtatttttt aaaacactaa 2760 aagcagcgtc actctaccta atgtctcact gttctgcaaa ggtggcaatg cttaaactaa 2820 ataatgaata aactgaatat tttggaaact gctaaattct atgttaaata ctgtgcagaa 2880 taatggaaac attacagttc ataataggta gtttggatat ttttgtactt gatttgatgt 2940 gacttttttt ggtataatgt ttaaatcatg tatgttatga tattgtttaa aattcagttt 3000 ttgtatcttg gggcaagact gcaaactttt ttatatcttt tggttattct aagccctttg 3060 ccatcaatga tcatatcaat tggcagtgac tttgtataga gaatttaagt agaaaagttg 3120 cagatgtatt gactgtacca cagacacaat atgtatgctt tttacctagc tggtagcata 3180 aataaaactg aatctcaaca taaaaaaaaa aaaaaaaaaa 3220

Claims (10)

A method for diagnosing gastric cancer by analyzing nucleic acids isolated from formalin-fixed paraffin-embedded (FFPE) tissue samples of a subject to be diagnosed,
The isolated nucleic acids are used to express human epidermal growth factor receptor (HER2), CCNE1 (cyclin E1), MYC (v-myc avian myelocytomatosis viral oncogene homolog), EGFR (epidermal growth factor receptor) and KRAS (Kirsten rat sarcoma viral oncogene performing a copy number variation (CNV) analysis of the homolog gene; And
Sequencing of the separated nucleic acid is performed to detect the presence of TP53 (tumor protein 53), APC (adenomatous polyposis coli), PIK3CA (phosphatidylinositol 4,5-bisphosphate 3-kinase catalytic subunit alpha), KRAS (Kirsten rat sarcoma viral oncogene homolog ), SMO (smoothened), STK11 (serine-threonine kinase 11), CDKN2A (cyclin-dependent kinase inhibitor 2A) and SMAD4 (SMAD family member 4)
Wherein CNV is present in at least one gene among the CNV analysis genes or when a mutation is present in at least one gene among the mutation determination genes, it is judged to be a gastric cancer.
delete 2. The method according to claim 1, wherein the information providing method comprises the step of selecting one or more selected from among SEQ ID NOS: 6 to 8 as the target sequence, and CCNE1 as the target sequence selected from SEQ ID NOS: 9 to 11, MYC is a target sequence selected from SEQ ID NO: 12 to SEQ ID NO: 14, EGFR is a target sequence selected from SEQ ID NO: 15 to SEQ ID NO: 17, and KRAS Is a target sequence selected from at least one selected from SEQ ID NO: 18 to SEQ ID NO: 20.
The information providing method according to claim 1, wherein the CNV is expressed in the following numerical range:
The gene CNV range of the first sequence is 5-65 copies;
A gene CNV range of SEQ ID NO: 2 sequence of 5-25 copies;
10-40 copies of the gene CNV range of the sequence of the third sequence;
The gene CNV range of the fourth sequence is 1-10 copies; And
The gene CNV range of SEQ ID NO: 5 sequence is 10-25 copies.
The method of claim 1, wherein the gastric cancer is gastric adenocarcinoma.
The method of claim 1, wherein the CNV analysis is performed using a nanostring nCounter analysis system.
delete 2. The method of claim 1, wherein the sequencing is semiconductor-based sequencing.
delete (Human epidermal growth factor receptor 2) using the nucleic acid isolated from formalin-fixed paraffin-embedded (FFPE) tissue samples of a subject to be diagnosed, , CCNE1 (cyclin E1), MYC (v-myc avian myelocytomatosis viral oncogene homolog), EGFR (epidermal growth factor receptor) and KRAS (Kirsten rat sarcoma viral oncogene homolog)
Sequencing of the separated nucleic acid is performed to detect the presence of TP53 (tumor protein 53), APC (adenomatous polyposis coli), PIK3CA (phosphatidylinositol 4,5-bisphosphate 3-kinase catalytic subunit alpha), KRAS (Kirsten rat sarcoma viral oncogene homolog ), SMO (smoothened), STK11 (serine-threonine kinase 11), CDKN2A (cyclin-dependent kinase inhibitor 2A) and SMAD4 (SMAD family member 4)
Wherein the CNV analysis gene is judged to be a gastric cancer when CNV appears in at least one gene of the CNV analysis gene or when a mutation appears in at least one gene of the mutation determination gene.

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