【摘要】 大肠杆菌的rnc基因编码产物为RNaseIII酶,RNaseIII酶能降解细菌中绝大多数dsRNA。利用来源于λ噬菌体的Red重组系统和重叠延伸PCR技术(gene splicing by overlap extension PCR,SOE-PCR),敲除了大肠杆菌origami(DE3)菌株的rnc基因,获得了RNaseIII缺失型菌株M-origami。利用电激法,将构建的TMV运动蛋白基因(movement protein gene,MP)的dsRNA表达载体LMP480导入M-origami菌株中,IPTG诱导表达的结果显示:构建的M-origami/LMP480原核表达系统能高效表达TMV运动蛋白基因的dsRNA。初步的抗病性鉴定显示,表达的dsRNA能够诱发烟草对TMV的抗性。更多还原

【Abstract】 The rncgene of E.colican encode the RNaseIII protein which can degrade most of dsRNA transcripts in E.coli.Thernc gene of origami(DE3) strain usingλ-dependent Red-mediated recombination systemand gene splicing by overlap extension PCRand the M-origamistrain deficient for RNaseIII.wat And achieved the vector of LMP480 that can express TMV MP dsRNA transcripts was amstueted and transformed it intoM-origamistrain by electroporation.Induced by IPTG,M-origami/LMP480 strains couldeffectively express TMV MP dsRNA transcriptsand the dsRNA could protect tobacco from the infection of TMV.更多还原