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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
Construction of transposition insertion library of Bacillus subtilis by using transposon TnYLB-1
【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。
【Abstract】 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.
【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;
【Key words】 Bacillus subtilis OKB105; transposon TnYLB-1; plasmid pMarA; transposable element Himar1; random mutagenesis;
【Key words】 Bacillus subtilis OKB105; transposon TnYLB-1; plasmid pMarA; transposable element Himar1; random mutagenesis;
【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)
- 【文献出处】 南京农业大学学报 ,Journal of Nanjing Agricultural University , 编辑部邮箱 ,2011年06期
- 【分类号】S476.1
- 【被引频次】30
- 【下载频次】795