China ; epidemiology ; Humans ; Yersinia Infections ; epidemiology ; microbiology ; Yersinia enterocolitica ; classification ; drug effects ; isolation & purification ; pathogenicity

Yersinia pseudotuberculosis is known to cause septicemia, mesenteric lymphadenitis enteritis and erythema nodosum. Most of the infections were found in European countries, but none in Korea ti11 now. For the first time in Korea Y. pseudotuberculosis was isolated form a 51-year-old ma1e with liver cirrhosis. The patient showed chills, abdominal pain and diarrhea followed by a comatose state. The organism was isolated from both blood and peritoneal fluid. The isolation and identification were difficult as the organism grew slowly and many of the characteristics were similar to other enteric bacilli. The isolate was susceptible to all antibiotics tested in vitro, but our chemotherapy with ampicillin and kanamycin did not save the patient's life.
Antibiotics/pharmacology ; Human ; Male ; Middle Age ; Septicemia/microbiology* ; Yersinia/drug effects ; Yersinia/isolation & purification ; Yersinia Infections/microbiology* ; Yersinia pseudotuberculosis Infections/microbiology*

OBJECTIVEThis study is to verify the use of rich BHI medium to substitute synthetic media for gene regulation studies in Yersinia pestis.

METHODSThe 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.

RESULTSWhen 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.

CONCLUSIONThe 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.

Bacterial Proteins ; genetics ; metabolism ; Bacteriological Techniques ; Culture Media ; pharmacology ; Gene Expression Regulation, Bacterial ; drug effects ; physiology ; Yersinia pestis ; metabolism ; physiology

To investigate molecular mechanism of traditional Chinese medicine Rheum offcinale against Yersinia pestis, whole genome DNA microarray that contains 4005 annotated genes of Y. pestis was used. The minimal inhibition concentration (MIC) of R. offcinale extract against Y. pestis was determined by liquid dilution method. The gene expression profile of Y. pestis was performed after exposured to R. offcinale extract at a concentration of 10 X MIC for 30 and 60 minutes. The total RNA extracted and purified from Y. pestis were reverse-transcribed to cDNA and labeled by Cy3-Cy5 dye. The labeled probes were hybridized to the microarray and the results were obtained by a laser scanner and analyzed by the SAM software. The microarray data was confirmed by RT-PCR. The platform of the DNA microarray-based bacteria transcriptional profiling was eshtablished. The results revealed general gene expression changes of Y. pestis were a global phenomenon. Down-regulation of genes encoding proteins involved in ribosome protein synthesis was a remarkable change. Genes encoding cell envelope and transport/binding proteins were the major changed genes of the Y. pestis in response to R. offcinale.
Bacterial Proteins ; genetics ; Down-Regulation ; Drugs, Chinese Herbal ; pharmacology ; Gene Expression Profiling ; Microbial Sensitivity Tests ; Oligonucleotide Array Sequence Analysis ; RNA, Bacterial ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Rheum ; chemistry ; genetics ; Yersinia pestis ; drug effects