枯草芽胞杆菌9407TnY LB-1转座子突变体库的构建
China Sciencepaper(2016)
Abstract
枯草芽胞杆菌9407(B9407)是1株对苹果轮纹病具有显著防效的生防菌株。为深入研究该菌株防病的分子机制,利用携带转座子T nY L B‐1的温敏型质粒pM arA构建了B9407的突变体库。通过电击转化将pM arA转化至B9407,经过30℃培养,37℃转座诱变,50℃进行pM arA质粒消除,最后通过抗性筛选,获得3500个转座子插入突变的单克隆,构建了B9407的突变体库。该转座子TnYLB‐1的转座效率达到2.4×10-3,pMarA的质粒消除率达97.3%。随机挑选12株突变子进行PCR及Southern blotting杂交,结果表明:转座子TnYLB‐1能够随机插入到B9407基因组上,大于90%的突变子为转座子单拷贝插入。B9407 T nY L B‐1转座子突变体库的构建为筛选和研究影响该菌株生防功能相关的基因并揭示其生防机制奠定了基础。
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Key words
plant pathology,Bacillus subtilis 9407,TnYLB-1,pMarA,transposon mutation,mutant library
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