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利用转座子TnYLB-1构建枯草芽孢杆菌的突变体文库 被引量:16

Construction of transposition insertion library of Bacillus subtilis by using transposon TnYLB-1
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摘要 为了鉴定枯草芽孢杆菌定殖和促进植物生长相关基因,用携带转座子TnYLB-1的穿梭载体pMarA转化枯草芽孢杆菌OKB105菌株。高温诱变条件下,筛选了2 000个转座子插入突变的单克隆,构建了OKB105菌株的突变体文库。对随机挑选的15个突变体采用PCR及Southern杂交验证,结果表明:转座子TnYLB-1以单拷贝、随机方式插入到OKB105菌株的染色体上。 In order to identify new genes involved in colonizing in plant rhizosphere and promoting plant growth of Bacillus subtilis,insertional mutagenesis of B.subtilis OKB105 was performed by use of a Himar1-based transposon TnYLB-1.The plasmid pMarA carrying the transposon TnYLB-1 were transformed into OKB105 and 2 000 transposants were selected randomly under the condition of high temperature.PCR and Southern hybridization analyses of 15 transposants showed that TnYLB-1 transposed into the B.subtilis chromosome randomly.
出处 《南京农业大学学报》 CAS CSCD 北大核心 2011年第6期77-81,共5页 Journal of Nanjing Agricultural University
基金 国家转基因生物新品种培育重大专项(2011ZX08004-004) 国际科技合作项目(2009DFA32740) 公益性行业(农业)科研专项(3-23) 国家863计划项目(2006AA10Z172) 国家大学生创新性实验计划项目(091030707)
关键词 枯草芽孢杆菌OKB105 TnYLB-1转座子 pMarA质粒 Himar1转座元件 随机诱变 Bacillus subtilis OKB105 transposon TnYLB-1 plasmid pMarA transposable element Himar1 random mutagenesis
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