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    Recombinant DNA Technology

    Uploaded by

    Monika Kanojia
    Recombinant DNA technology involves introducing foreign DNA into an organism's genome. It has multiple steps and tools, including restriction enzymes to cut DNA, vectors to carry DNA into a host, and DNA ligase to join DNA strands. The process includes isolating DNA, cutting it with enzymes, amplifying copies with PCR, ligating the DNA into a vector, and transforming the recombinant DNA into a host organism. Applications include producing medicines, genetically modifying crops, and gene therapy.

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    Recombinant DNA Technology

    Uploaded by

    Monika Kanojia
    78 views5 pages
    Recombinant DNA technology involves introducing foreign DNA into an organism's genome. It has multiple steps and tools, including restriction enzymes to cut DNA, vectors to carry DNA into a host, and DNA ligase to join DNA strands. The process includes isolating DNA, cutting it with enzymes, amplifying copies with PCR, ligating the DNA into a vector, and transforming the recombinant DNA into a host organism. Applications include producing medicines, genetically modifying crops, and gene therapy.

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    Recombinant DNA technology involves introducing foreign DNA into an organism's genome. It has multiple steps and tools, including restriction enzymes to cut DNA, vectors to carry DNA into a host, and DNA ligase to join DNA strands. The process includes isolating DNA, cutting it with enzymes, amplifying copies with PCR, ligating the DNA into a vector, and transforming the recombinant DNA into a host organism. Applications include producing medicines, genetically modifying crops, and gene therapy.

    Copyright:

    © All Rights Reserved
    Download as DOCX, PDF, TXT or read online from Scribd
    Download as docx, pdf, or txt
    78 views5 pages

    Recombinant DNA Technology

    Uploaded by

    Monika Kanojia
    Recombinant DNA technology involves introducing foreign DNA into an organism's genome. It has multiple steps and tools, including restriction enzymes to cut DNA, vectors to carry DNA into a host, and DNA ligase to join DNA strands. The process includes isolating DNA, cutting it with enzymes, amplifying copies with PCR, ligating the DNA into a vector, and transforming the recombinant DNA into a host organism. Applications include producing medicines, genetically modifying crops, and gene therapy.

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    © All Rights Reserved
    Download as DOCX, PDF, TXT or read online from Scribd
    Download as docx, pdf, or txt
    of 5

    A technique mainly used to change the phenotype of an organism (host) when a genetically

    altered vector is introduced and integrated into the genome of the organism. So, basically, this
    process involves the introduction of a foreign piece of DNA structure into the genome which
    contains our gene of interest. This gene which is introduced is the recombinant gene and the
    technique is called the recombinant DNA technology.

    There are multiple steps, tools and other specific procedures followed in the recombinant
    DNA  technology, which is used for producing artificial DNA to generate the desired product.
    Let’s understand each step more in detail.

    Table of Contents

    ● Explanation
    ● Tools
    ● Process
    ● Application
    ● DNA Cloning
    ● Applications Of  Gene Cloning

    What is Recombinant DNA Technology?


    The technology used for producing artificial DNA through the combination of different genetic
    materials (DNA) from different sources is referred to as Recombinant DNA
    Technology. Recombinant DNA technology is popularly known as genetic engineering. 

    The recombinant DNA technology emerged with the discovery of restriction enzymes in the year
    1968 by Swiss microbiologist Werner Arber,

    Inserting the desired gene into the genome of the host is not as easy as it sounds. It involves the
    selection of the desired gene for administration into the host followed by a selection of the
    perfect vector with which the gene has to be integrated and recombinant DNA formed.

    Thus the recombinant DNA has to be introduced into the host. And at last, it has to be
    maintained in the host and carried forward to the offspring.

    Also Refer- Genes
    Tools Of Recombinant DNA Technology
    The enzymes which include the restriction enzymes help to cut, the polymerases- help to
    synthesize and the ligases- help to bind. The restriction enzymes used in recombinant DNA
    technology play a major role in determining the location at which the desired gene is inserted into
    the vector genome. They are two types, namely Endonucleases and Exonucleases.

    The Endonucleases cut within the DNA strand whereas the Exonucleases remove the nucleotides
    from the ends of the strands. The restriction endonucleases are sequence-specific which are
    usually palindrome sequences and cut the DNA at specific points. They scrutinize the length of
    DNA and make the cut at the specific site called the restriction site. This gives rise to sticky ends
    in the sequence. The desired genes and the vectors are cut by the same restriction enzymes to
    obtain the complementary sticky notes, thus making the work of the ligases easy to bind the
    desired gene to the vector.

    The vectors – help in carrying and integrating the desired gene. These form a very important part
    of the tools of recombinant DNA technology as they are the ultimate vehicles that carry forward
    the desired gene into the host organism. Plasmids and bacteriophages are the most common
    vectors in recombinant DNA technology that are used as they have a very high copy number. The
    vectors are made up of an origin of replication- This is a sequence of nucleotides from where the
    replication starts, a selectable marker – constitute genes which show resistance to certain
    antibiotics like ampicillin; and cloning sites – the sites recognized by the restriction enzymes
    where desired DNAs are inserted.

    Host organism – into which the recombinant DNA is introduced. The host is the ultimate tool of
    recombinant DNA technology which takes in the vector engineered with the desired DNA with the
    help of the enzymes.
    There are a number of ways in which these recombinant DNAs are inserted into the host, namely
    – microinjection, biolistics or gene gun, alternate cooling and heating, use of calcium ions, etc.

    Also Read: Bioinformatics 

    Process of Recombinant DNA Technology


    The complete process of recombinant DNA technology includes multiple steps, maintained in a
    specific sequence to generate the desired product.

    Step-1. Isolation of Genetic Material.

    The first and the initial step in Recombinant DNA technology is to isolate the desired DNA in its
    pure form i.e. free from other macromolecules.

    Step-2.Cutting the gene at the recognition sites.

    The restriction enzymes play a major role in determining the location at which the desired gene is
    inserted into the vector genome. These reactions are called ‘restriction enzyme digestions’.

    Step-3. Amplifying the gene copies through Polymerase chain reaction (PCR).

    It is a process to amplify a single copy of DNA into thousands to millions of copies once the
    proper gene of interest has been cut using restriction enzymes.

    Step-4. Ligation of DNA Molecules.

    In this step of Ligation, the joining of the two pieces – a cut fragment of DNA and the vector
    together with the help of the enzyme DNA ligase.

    Step-5. Insertion of Recombinant DNA Into Host.

    In this step, the recombinant DNA is introduced into a recipient host cell. This process is termed
    as Transformation. Once the recombinant DNA is inserted into the host cell, it gets multiplied
    and is expressed in the form of the manufactured protein under optimal conditions.

    As mentioned in Tools of recombinant DNA technology, there are various ways in which this can
    be achieved. The effectively transformed cells/organisms carry forward the recombinant gene to
    the offspring.

    Also Read: R-Factor

    Application of Recombinant DNA Technology


    ● DNA technology is also used to detect the presence of HIV in a person.
    ● Gene Therapy – It is used as an attempt to correct the gene defects which give rise to
    heredity diseases.
    ● Clinical diagnosis – ELISA is an example where the application of recombinant
    ● Recombinant DNA technology is widely used in Agriculture to produce genetically-
    modified organisms such as  Flavr Savr tomatoes, golden rice rich in proteins, and Bt-
    cotton to protect the plant against ball worms and a lot more.
    ● In the field of medicines, Recombinant DNA technology is used for the production of
    Insulin.
    Also Refer: Genetically Modified Organisms (GMO)
    DNA Cloning
    A clone is a cluster of individual entities or cells that are descended from one progenitor. Clones
    are genetically identical as the cell simply replicates producing identical daughter cells every
    time. Scientists are able to generate multiple copies of a single fragment of DNA, a gene which
    can be used to create identical copies constituting a DNA clone. DNA cloning takes place
    through the insertion of DNA fragments into a tiny DNA molecule. This molecule is made to
    replicate within a living cell, for instance, a bacterium. The tiny replicating molecule is known as
    the carrier of the DNA vector.

    Yeast cells, viruses, and Plasmids are the most commonly used vectors. Plasmids are circular
    DNA molecules that are introduced from bacteria. They are not part of the main cellular genome.
    It carries genes, which provide the host cell with beneficial properties such as mating ability, and
    drug resistance. They can be conveniently manipulated as they are small enough and they are
    capable of carrying extra DNA which is weaved into them.

    Explore more: Genetic Disorders.

    Applications Of  Gene Cloning


    Listed below are the applications of gene cloning:

    ● Gene Cloning plays an important role in the medicinal field. It is used in the production of
    hormones, vitamins and antibiotics.
    ● Gene cloning finds its applications in the agricultural field. Nitrogen fixation is carried out
    by cyanobacteria wherein desired genes can be used to enhance the productivity of
    crops and improvement of health. This practice reduces the use of fertilizers hence
    chemical-free produce is generated
    ● It can be applied to the science of identifying and detecting a clone containing a
    particular gene which can be manipulated by growing in a controlled environment
    ● It is used in gene therapy where a faulty gene is replaced by the insertion of a healthy
    gene. Medical ailments such as leukaemia and sickle cell anaemia can be treated with
    this principle.
    Also Refer- Gene Therapy.

    Stay tuned with BYJU’S to learn more about the Recombinant DNA Technology, its tools,
    procedure and other related topics at  BYJU’S Biology.

    Frequently Asked Questions

    Explain the roles of the following:


     
    (a) Restriction Enzymes
     
    (b) Plasmids
    a) Restriction enzymes are molecular scissors used in molecular biology for cutting DNA
    sequences at a specific site. It plays an important role in gene manipulation.
     
    b) Plasmid is an extra-chromosomal DNA molecule in bacteria that is capable of replicating,
    independent of chromosomal DNA. They serve as a vehicle to carry a foreign DNA sequence into
    a given host cell.

    Explain PCR.
    PCR stands for Polymerase Chain Reaction, a method of amplifying fragments of DNA. This
    method can make multiple copies of even a single DNA fragment or the gene of interest, in a test
    tube.

    Discuss the applications of recombination from the point of view of genetic


    engineering.
    Applications of recombinant DNA in genetic engineering are:

    1. For the production of vaccines like the hepatitis B vaccine.


    2. Production of transgenic plants with improved qualities like insect and drought
    resistance and nutritional enrichment.
    3. Therapeutic protein production like insulin.
    4. Gene therapy in diseases like cancer, SCID etc.
    5. Production of transgenic animals with improved quality of milk and egg.

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