Construction of a MluI-YAC library from barley (Hordeum vulgare L.) and analysis of YAC insert terminal regions.
作者： Michael Kleine , Christian Jung , Wolfgang Michalek , Thomas Diefenthal , Harald Dargatz
DOI: 10.1139/G97-116
关键词:
摘要: We describe the construction of a specific yeast artificial chromosome (YAC) library from barley (Hordeum vulgare L.) using vector pYAC-RC. The was generated by total digestion high molecular weight DNA with infrequently cutting restriction enzyme MluI. Only 10-30% colonies were recombinant, as visualized red-white selection and subsequent pulsed-field gel electrophoresis analysis. About 17 000 individual recombinant YAC clones insert sizes ranging 50 to 700 kb, mean 170 selected. No chloroplast sequences detected proportion containing BARE-1 copia-like retroelements is about 5%. Screening single-copy RFLP marker closely linked Mla locus yielded three identical same size. Insert termini randomly chosen investigated respect their redundancy in genome compared an EcoRI-based library, resulting fourfold enrichment at MluI vector-insert junctions.
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