大肠埃希菌sdiA基因缺失株的建立  

Construction of The sdiA Gene Mutant in Escherichia coli Strains

作　　者：伍彬宁 蔡壬辛[1] 曾建明[1] 鄂顺梅[1] 李有强[1] 叶大柠 鲁洋[1] 陈茶[1] 

机构地区：[1]广州中医药大学第二临床医学院检验医学部,广东广州510006

出　　处：《国际检验医学杂志》2015年第17期2466-2468,共3页International Journal of Laboratory Medicine

基　　金：国家自然科学基金资助项目(81071397;81271909);广东省自然科学基金资助项目(S2013010012970)

摘　　要：目的为研究HSL信号分子对大肠埃希氏菌的影响而建立大肠埃希菌sdiA基因缺失株。方法通过Red重组系统敲除大肠杆菌BW25113和SM10λpir的sdiA基因,PCR测序鉴定;不同时间点检测A600值,绘制细菌生长曲线;荧光定量PCR检测csgD、csgB基因水平。结果 PCR测序结果经DNAman软件比对分析,与预想结果一致;与野生株相比,sdiA基因突变对细菌生长曲线无明显影响。结论成功建立BW25113、SM10λpir的sdiA基因缺失株,为后续研究铜绿假单胞菌信号分子HSL对大肠埃希菌的影响奠定了基础。Objective To build sdiA gene muants of Escherichia coli BW25113 and SM10λpir strains .Methods The sdiA gene of Escherichia coli BW25113 and SM10λpir were knocked out with Red recombination system .The mutants were detected by PCR technique ,and the following sequencing results were analyzed through alignment by software DNAman .Growth curve of strains were assayed by detecting the A600 value at different time point .The csgD and csgB genes expression levels were detected by fluo‐rescent quantitative PCR in SM10λpir sdiA mutant thereafter .Results PCR products electrophoresis showed a shorten fragment in mutants comparing to their parental strains BW25113 and SM10λpir ,and it was confirmed by DNA sequencing .The growth curves and cells morphology in Gram stain were similar in both mutants and wild‐type strains .The expression of csgD and csgB genes was higher in SM10λpir sdiA mutants than those in wild‐type strain .Conclusion The mutants of sdiA deletion of Escherichia coli BW25113 and SM10λpir were constructed successfully ,and it will be the foundation for further research of the effects of the signal molecular HSL on Escherichia coli .

关 键 词：大肠埃希菌 基因敲除 RED重组系统 sdiA基因 HSL信号分子 接合反应 

分 类 号：R440[医药卫生—诊断学]