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ABSTRACT: In order to develop a detailed physical map of the thermo-sensitive genie male-sterile (TGMS) gene-encompassing region and
finally clone the TGMS gene, a high-quality rice bacterial artificial chromosome (BAC) library from TGMS rice 5460S was constructed.
The method of constructing BAC library was examined and optimized. The 5460S library consists of 19 584 BAC clones with an
average insert size of 110 kb, which represents about 5 times rice haploid genome equivalents. Rice inserts of up to 140 kb
and 250 kb were isolated and appeared stable after 100 generations of serial growth. Hybridization of BAC clones with mitochondrial
and chloroplastic genes as probes demonstrated that this library has no organellar contamination. The 5460S library was screened
with 3 molecular markers linked totms 1 gene as probes and at least 1 BAC clone was identified with each probe. The insert ends of positive clones were successfully
isolated using thermal asymmetric interlaced PCR (TAIL-PCR) technique.
Science in China Series C Life Sciences 11/1999; 42(6):599-606. · 1.61 Impact Factor