Construction and characterization of a bacterial artificial chromosome library of rice 5460F

Thermo-sensitive genic male sterile (TGMS) rice has a number of desirable c haracteristics for hybrid rice production. Many studies have demonstrated t hat the sterility of TGMS rice is controlled by a single recessive gene. It has been mapped for the first time on chromosome 8 and named tms1. Several AFLP markers which tightly linked to the tms1 gene have been identified re cently. In order to develop a detailed physical map of the tms1 gene-encomp assing region and finally clone the tmsl gene, a bacterial artificial chrom osome (BAC) library of rice 5460F (the fertile mutant line of TGMS rice 546 0S) using a modified vector pECBAC1 has been constructed. The constructed 5 460F library consists of 16 896 clones with an average insert size of 119 k b, which represents about 4.7 times rice haploid genome equivalents. Neithe r chloroplast nor mitochondrial DNA was detected from the library. The libr ary was screened with three single copy sequence amplified fragment length polymorphism (AFLP) markers which tightly linked to tmsl gene as probes and eight positive clones were identified.