【摘要】 目的建立信号标签诱变突变体穿梭质粒,为构建信号标签诱变突变体文库及病原微生物毒力基因筛选打下基础。方法通过PCR扩增信号标签(signature tags,STs)片段,导入载体pC6,构建重组质粒pC6RS,转化到大肠杆菌CC118中,提取重组质粒后转化到大肠杆菌S17-1中。结果经过PCR、核苷酸序列测定鉴定,成功地构建了pC6RS质粒。更多还原

【Abstract】 Objective To construct the shuttle plasmid of Signature-Tagged Mutagenesis technology and lay the basis to establish the mutant library and virulence genes selection of pathogenic microorganism.Methods The DNA signature tags was obtained by PCR from the single stands DNA tags.The signature tags was cloned into pC6 to construct recombinant plasmid pC6RS.Then it was transformed into E.coli CC118 to produce multiple recombinant plasmid.pC6RS extracted from CC118 was subsequently transformed into E.coli S17-1. Results The recombinant plasmid pC6RS identified by enzyme digestion and DNA sequencing was successfully constructed.更多还原