Papers by Remedios Crespillo
Abstract. Establishment of protoplast techniques for conifers is of crucial importance to forest ... more Abstract. Establishment of protoplast techniques for conifers is of crucial importance to forest biotechnology. Protoplasts can be used to regenerate whole trees and to study transcriptional regulation in woody plants by promoter analysis. Here we describe a tech-nique for isolating protoplasts from Pinus pinaster shoots. Protoplast viability was deter-mined by fluorescein diacetate staining. Transient gene expression was studied after electroporation of protoplasts with plasmid DNA containing a b-glucuronidase (GUS) re-porter gene. The 35S promoter and a pine (cytosolic glutamine synthetase) promoter both were able to drive GUS expression, indicating applicability of conifer protoplasts as a model to study gene promoters using transient expression. Isolation of gymnosperm shoot protoplasts Gómez-Maldonado et al.
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Avances En El Metabolismo Del Nitrogeno Bioquimica Fisiologia Y Biologia Molecular 2000 Isbn 84 7496 842 9 Pags 225 232, 2000
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Avances En El Metabolismo Del Nitrogeno De La Fisiologia a La Biologia Molecular Iv Reunion Nacional Marbella 1997 1998 Isbn 84 472 0402 2 Pags 535 542, 1998
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Tolerance to ammonium nutrition in plants can be related to their ability to detoxify ammonium vi... more Tolerance to ammonium nutrition in plants can be related to their ability to detoxify ammonium via nitrogen assimilation in roots. Here, we report that sorghum–sudangrass (Sorghum bicolor L. S. bicolor var. sudanense) hybrids exhibited enhanced biomass production under high levels of inorganic nitrogen supply as well as increased capacity for nitrogen assimilation in roots. Glutamine synthetase (GS, EC 6.3.1.2) activity and protein accumulated in roots at increasing concentrations of either nitrate or ammonium, with particularly high levels of GS in ammonium-treated plants. Ammonium but not nitrate differentially regulated two distinct cytosolic GS (GS1) isoforms composed by polypeptides of similar size but different charge. The comparative analysis of GS gene sequences and the deduced GS1 polypeptides suggested that the two GS1 isoforms were the expression products of SbGln1.2 and SbGln1.3 genes. SbGln1.3 expression was shown to be upregulated by high levels of inorganic nitrogen s...
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Somatic Cell Genetics and Molecular Genetics of Trees, 1996
MOLECULAR ANALYSIS OF PINE FERREDOXIN-DEPENDENT GLUTAMATE SYNTHASE ANGEL GARCIA-GUTIERREZ, FERNAN... more MOLECULAR ANALYSIS OF PINE FERREDOXIN-DEPENDENT GLUTAMATE SYNTHASE ANGEL GARCIA-GUTIERREZ, FERNANDO GALLARDO, FRANCISCO R. CANTON, REMEDIOS CRESPILLO AND FRANCISCO M. CANOVAS Laboratorio de Bioquimica y Biologia Molecular, ...
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In a previous work we reported the molecular characterization of a glutamine synthetase (GS; EC 6... more In a previous work we reported the molecular characterization of a glutamine synthetase (GS; EC 6.3.1.2.) complementary DNA from a woody plant (Cantbn et al. (1993) Plant Mol. Biol. 22, 819-828). The isolated cDNA (pGSP114) encoding a Scots pine (Pinus sylvestris) cytosolic subunit, has been subcloned into the expression vector pET3c to overproduce the GS polypeptide in Escherichia coli cells. The recombinant GS protein showed the same molecular size as a native Scots pine GS subunit. Antibodies against the pET3c-GSPll4 encoded protein were raised in rabbits by injecting purified preparations and specificity was determined by immunoprecipitation of GS activity present in pine crude extracts. In spite of the antibodies were able to recognize both cytosolic and chloroplastic GS in tomato plants, they were unable to immunodetect chloroplastic GS in green cotyledons of pine seedlings and cytosolic GS was the unique recognized polypeptide. Unlike to that found in other plant species, cyt...
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Plant Physiology and Biochemistry, 1998
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Physiologia Plantarum, 1998
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FEBS Letters, 1996
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High-level expression of Pinus sylvestris glutamine synthetase in
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Papers by Remedios Crespillo