大肠杆菌多重耐药调控基因soxS的克隆及其原核表达  

Cloning and prokaryotic expression of muti-drug resistance regulating gene soxS of Escherichia coli

作　　者：于晓颖[1] 刘玉堂[1] 丛薇[1] 刘树明[1] 马红霞[1] 

机构地区：[1]吉林农业大学动物科学技术学院,吉林长春130118

出　　处：《中国兽医学报》2010年第10期1318-1320,1325,共4页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(30400326);吉林省科技厅项目(20010538;20030425;20050124);吉林省教育厅项目(2005042)

摘　　要：根据GenBank中大肠杆菌的soxS基因序列设计引物,以大肠杆菌ATCC25922的基因组为模板,PCR扩增出约324 bp的soxS基因片段,将所得片段与pMD18-Tsi mple vector连接,转化至J M109大肠杆菌中,成功地筛选到阳性克隆,其质粒序列测序结果与GenBank中报道一致。从阳性克隆中提取质粒,经BamHⅠ和XhoⅠ酶解,回收324 bp的目的片段,定向克隆到pET-28a表达载体中,提取质粒,再次转化到大肠杆菌BL21(DE3)中,成功地筛选出阳性克隆。经IPTG诱导阳性菌,获得表达产物,通过SDS-PAGE检测出soxS基因的表达。A constructed soxS-pMD-18T was generated by inserting the sequence of 324 bp obtained by a PCR into pMD18-T simple vector and selecting the sense clones,The result showed that the cloned sequence coincides with the designed sequence by sequencing.This construction was digested with the same enzyme（BamH Ⅰ and Xho Ⅰ） and ligated the pET-28a vector.Then the plasmid soxS-pET-28a was transformed to the competent cell of BL21（DE3）.The sense clone was induced with IPTG.The expression of soxS was observed on SDS-PAGE.

关 键 词：大肠杆菌 soxS基因 克隆 原核表达 

分 类 号：S852.6[农业科学—基础兽医学]