Detection, isolation, and characterization of minisatellite sequences for fingerprinting analysis in Phaseolus
1994
Gonzalez, Delkin Orlando | Tenjo F, Fernando Alberto | Tohme M, Joseph
Minisatellites are a short core of consensus sequence repeated in tandem and variable in number along the genome of an individual. Multilocus DNA fingerprinting is a powerful tool for germplasm characterization, evolutionary and population studies. The determination of such sequences has already been identified in different plant species. In order to detect and isolate minisatellites sequences in beans two Phaseolus vulgaris and one Phaseolus acutifolius genomic libraries were constructed in lambda NM1149. Human minisatellites and an internal sequence from bacteriophage M13 were used as probes to detect minisatellites in beans. After purification with M13, 125 possible positive clones were isolated but only 28 clones were selected and purified. They were used as probes in DNA fingerprinting of three different Phaseolus species. One clone of 6Kb in size is useful in detecting polymorphism among individuals from interspecific crosses and another one of 2 Kb in size seems to be highly specific to Phaseolus species. We plan to further characterize the positive clones and use them to detect possible species-specific markers for genome and interspecific crosses analyses.
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