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

作　　者：马欣[1] 刘俊[1] 乔俊卿[1] 伍辉军[1] 高学文[1] 

机构地区：[1]南京农业大学植物保护学院/农作物生物灾害综合治理教育部重点实验室,江苏南京210095

出　　处：《南京农业大学学报》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)

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.

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 

分 类 号：S432.43[农业科学—植物病理学]