霍乱弧菌溶原性噬菌体CTXΦ的氯霉素抗性基因标记及诱导  

Chloramphenicol resistance marker exchange of lysogenic phage CTXΦ of Vibrio cholerae and its induction

作　　者：刘广文[1] 闫梅英[1] 祁国明[1] 刘延清[1] 高守一[1] 阚飙[1] 

机构地区：[1]中国疾病预防控制中心传染病预防控制所卫生部医学分子细菌学重点实验室,北京102206

出　　处：《中华微生物学和免疫学杂志》2004年第4期253-257,共5页Chinese Journal of Microbiology and Immunology

基　　金：国家重大基础研究项目 (973 )课题(G19990 5 410 2 );国家高技术研究发展计划 (863)资助项目 (2 0 0 1AA2 15 191 2 )

摘　　要：目的　对携带霍乱毒素基因的溶原性噬菌体CTXΦ进行遗传标记 ,以深入研究CTXΦ对霍乱弧菌的转染机制。方法　将CTXΦ基因组中ctxAB的上下游同源序列连接到cat基因的两侧 ,然后通过自杀质粒介导的DNA同源重组技术将霍乱菌株N16 96 1染色体上CTXΦ基因组中的ctxAB基因缺失掉 ,替换以氯霉素抗性基因 ,对CTXΦ基因组进行遗传标记 ,然后用丝裂霉素C诱导这种经抗性基因标记的噬菌体 ,再利用它对古典型霍乱弧菌的转染检测活性。结果　获得了ctxAB缺失、带有cat遗传标记的重组菌株N Φc;诱导出的CTXΦc噬菌体颗粒成功感染了霍乱菌株 1119。结论　N Φc中的CTXΦ带有了氯霉素抗性标记 。Objective To mark the CTXΦ genome with chloramphenicol acetyltransferase gene ( cat ) and to induce it as a tool to study the CTXΦ′s transduction in V.cholerae . Methods ctxAB gene of CTXΦ on chromosome V. cholerae N16961 was exchanged by cat gene, which was flanked with upstream and downstream fragment of ctxAB , through homologous recombination mediated by the suicide plasmid. The cat marked CTXΦ genome was induced with mitomycin C to test its transfection among classical type V. cholerae strains. Results cat gene replaced ctxAB which integrates on the chromosome, and this cat marked CTXΦ genome, N Φc, can be induced and infected the tested V. cholerae strain. Conclusion We obtained a cat marked CTXΦ and can be used to study the horizontal transfer mechanism of CTXΦ between different V. cholerae strains.

关 键 词：霍乱弧菌 溶原性噬菌体 氯霉素抗性基因 CTXΦ基因 DNA同源重组技术 

分 类 号：R346[医药卫生—基础医学]