Use of Rich BHI Medium Instead of Synthetic TMH Medium for Gene Regulation Study in Yersinia pestis

查看全文 作　　者：ZHANG Yi [1]Quan;MA Li [1,2]Zhi;WANG [1,3]Li;GAO [1,4]He;TAN Ya [1]Fang;GUO Zhao [1]Biao;QIU Jing [3]Fu;YANG Rui [1]Fu;ZHOU Dong [1]Sheng 机构地区：[1]State Key Laboratory of Pathogen and Biosecurity, Beo'ing Institute of Microbiology and Epidemiology, Beijing 100071, China;[2]Department of Emergency Medicine, General Hospital of Chinese People's Armed Police Forces, Beijing 100039, China;[3]Department of Medical Laboratory, Chongqing Medical University, Chongqing 400016, China,;[4]State Key Laboratory for Infectious Disease Prevention and Control National Institute for Communicable Disease Control and Prevention, Chinese Centre for Disease Control and Prevention, Beijing 102206, China 出　　处：《Biomedical and Environmental Sciences》索引2012年第25卷第6期,共6页 基　　金：supported by the National Natural Science Foundation of China (30930001 and 30900823) 摘　　要：Objective This study is to verify the use of rich BHI medium to substitute synthetic media for gene regulation studies in Yersinia pestis. Methods The transcriptional regulation of rovA by PhoP or via temperature upshift, and that of pla by CRP were investigated when Y. pestis was cultured in BHI. After cultivation under 26 °C, and with temperature shifting from 26 to 37 °C, the wild-type (WT) strain or its phoP or crp null mutant (ΔphoP or Δcrp, respectively) was subject to RNA isolation, and then the promoter activity of rovA or pla in the above strains was detected by the primer extension assay. The rovA promoter-proximal region was cloned into the pRW50 containing a promoterless lacZ gene. The recombinant LacZ reporter plasmid was transformed into WT and ΔphoP to measure the promoter activity of rovA in these two strains with the β-Galactosidase enzyme assay system. Results When Y. pestis was cultured in BHI, the transcription of rovA was inhibited by PhoP and upon temperature upshift while that of pla was stimulated by CRP. Conclusion The rich BHI medium without the need for modification to be introduced into the relevant stimulating conditions (which are essential to triggering relevant gene regulatory cascades), can be used in lieu of synthetic TMH media to cultivate Y. pestis for gene regulation studies. 关 键 词：鼠疫耶尔森氏菌 基因调控 合成 启动子活性 LACZ基因 转录调控 培养基 半乳糖苷酶