title: Use of rich BHI medium instead of synthetic TMH medium for gene regulation study in Yersinia pestis.
abstract: 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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, China.
authors.forename: Yi Quan
authors.initials: YQ
authors.lastname: Zhang
authors.forename: Li Zhi
authors.initials: LZ
authors.lastname: Ma
authors.forename: Li
authors.initials: L
authors.lastname: Wang
authors.forename: He
authors.initials: H
authors.lastname: Gao
authors.forename: Ya Fang
authors.initials: YF
authors.lastname: Tan
authors.forename: Zhao Biao
authors.initials: ZB
authors.lastname: Guo
authors.forename: Jing Fu
authors.initials: JF
authors.lastname: Qiu
authors.forename: Rui Fu
authors.initials: RF
authors.lastname: Yang
authors.forename: Dong Sheng
authors.initials: DS
authors.lastname: Zhou
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D001431:Bacteriological Techniques; D003470:Culture Media / Q000494:pharmacology*; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects / Q000502:physiology*; D015010:Yersinia pestis / Q000378:metabolism* / Q000502:physiology
chemical_list: D001426:Bacterial Proteins; D003470:Culture Media