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Construcción de una genoteca de cDNA de fríjol (Phaseolus vulgaris L.) para mapeo genético

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Abstract:

A small cDNA library from beans (Phaseolus vulgaris L.) of about 1500 clones was constructed, to further saturate the bean RFLP linkage map. The primarily synthesized cDNAs were amplified by PCR using the adaptors as primers for amplification (Jepson et al., 1991). Inserts in the range of 500 bp were obtained. 93 clones were singled for further analysis. They showed a degree of repeatibility of around 61 %. Around 80% of the unique clones were single copy, and 20% were low copy sequences as expected from a cDNA library. Three pairs of parental bean lines were chosen for their agronomical traits, and evaluated for polymorphism, which was highest as revealed by digestion with EcoRV (77%), followed by DraI (73%), EcoRI (63%) and HindIIl (60%). The highest polymorphism was observed between the pair DOR60 and APNI8, 71% for EcoRV and 57% for EcoRI, respectively. Two clones of the two groups with the most repeated clones were analyzed by slot blot hybridization against the other clones and ribosomal DNA, to understand the origin of the repetitions. Only one clone seemed to be of ribosomal origin, as confirmed by the patterns obtained by hybridization to bean genomic DNA digested with HaelIl, implying that the whole group to which it belonged was of ribosomal origin. It can be explained by the combined utilization of the PCR amplification methodology and the multipleprimers for the synthesis of the first cDNA strand.

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Plant pathogens and resistance mechanisms

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Volumen45
Issue1
Páginas7 - 13
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