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Construction and characterization of a bacterial artificial chromosome library of thermo-sensitive genic male-sterile rice 5460S
作者姓名:邱芳  金德敏  伏健民  张超良  谢纬武  王斌  杨仁崔  张洪斌
基金项目:Project supported by the Rockefeller Foundation and China National High-Tech Program.
摘    要: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 to tmsl 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.

收稿时间:14 April 1999

Construction and characterization of a bacterial artificial chromosome library of thermo-sensitive genie male-sterile rice 5460S
QIU FangJIN DeminFU JianminZHANG ChaoliangXIE WeiwuYANG RencuiZHANG HongbinWANG Bin.Construction and characterization of a bacterial artificial chromosome library of thermo-sensitive genic male-sterile rice 5460S[J].Science China Life sciences,1999,42(6):599-606.
Authors:QIU FangJIN DeminFU JianminZHANG ChaoliangXIE WeiwuYANG RencuiZHANG HongbinWANG Bin
Institution:(1) Institute of Genetics, Chinese Academy of Sciences, 100101 Beijing, China;(2) Fujian Agricultural University, 350002 Fuzhou, China;(3) Department of Soil and Crop Sciences, Texas A & M University, 77843 Texas, USA
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 mitochondria! and chloroplastic genes as probes demonstrated that this library has no organellar contamination. The 5460S library was screened with 3 molecular markers linked to tms 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.
Keywords:rice thermo-sensitive genie-male sterile gene  bacterial artificial chromosome library  thermal asymmetric interlaced PCR  
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