克雷伯氏肺炎桿菌中受血清抑制之cad及tdc基因區功能研究

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doi:10.6342/NTU.2008.00414

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克雷伯氏肺炎桿菌中受血清抑制之cad及tdc基因區功能研究

Functional analysis of serum suppressed genes cad and tdc region in Klebsiella pneumoniae

林蕙心 , Masters　　Advisor：王錦堂　　

繁體中文 DOI： 10.6342/NTU.2008.00414

克雷伯氏肺炎桿菌； cad 基因； tdc 基因；酸性環境存活；膽鹽壓力；體內競爭實驗； Klebsiella pneumoniae ； cad ； tdc ； acid tolerance response ； bile salt stress ； in vivo competition

Abstract │ Reference (57) │ Altmetrics

Abstract 〈TOP〉

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Community acquired primary pyogenic liver abscess with metastatic complications caused by Klebsiella pneumoniae is a global emerging infectious disease, however the pathogenesis mechanism remains unclear. To investigate the possible pathogenesis mechanism, in the previous study, we used health human serum to mimic the in vivo environment and used microarray to find novel genes associated with virulence. Results showed that the RNA expression level of cad region and tdc region in community acquired K. pneumoniae representative strain NTUH-K2044 were down-regulated under health human serum treatment. We focused on studying the genetic and functional characteristics of these two gene loci. Analyzing the transcriptional unit of cad region and tdc region, we found cad region was composed of known transcriptional activator gene cadC and cadBA operon; cadBA operon included yjdL and lysS. YjdL and LysS might involve in lysine metabolism together with protein gene products of cadBA. The transcriptional activator gene in tdc region, tdcA, was separately transcripted from other genes of tdc region, which is different from the tdc operon of E. coli. 
In functional study, the Lys decarboxylase activity of the constructed cad region gene mutants, including ΔcadC and ΔcadA mutants, analyzed by Moeller test was negative and the survival rate under acid environment was decreased. Also, the resistance to bile salt was decreased in ΔcadC and Δtdc mutants, suggesting cadC can increase the survival rate under acid and bile salt stress. Using microarray to compare genes expression of NTUH-K2044 wild type and Δcad mutant, we found expression level of tdc region was decreased in Δcad mutant. By RT-qPCR, we confirmed that RNA expression of tdc region was decreased in ΔcadC mutant. cadC might control tdc region expression. In in vivo virulence analysis, we used intraperitoneal inoculation and intragastric infection in BALB/c mice model to perform in vivo competition assay. The results revealed that colonization ability of Δcad and Δtdc mutants were both weaker than that of wild type in liver, spleen, colon and cecum. This suggested that cad and tdc gene region play important roles in in vivo colonization.

Reference ( 57 ) 〈TOP〉

Bearson, S., Bearson, B. and Foster, J.W. (1997) Acid stress responses in enterobacteria. FEMS Microbiol Lett, 147, 173-180.
連結：
Begley, M., Gahan, C.G. and Hill, C. (2005) The interaction between bacteria and bile. FEMS Microbiol Rev, 29, 625-651.
連結：
Bernstein, C., Bernstein, H., Payne, C.M., Beard, S.E. and Schneider, J. (1999) Bile salt activation of stress response promoters in Escherichia coli. Curr Microbiol, 39, 68-72.
連結：
Bobrov, A.G. and Perry, R.D. (2006) Yersinia pestis lacZ expresses a beta-galactosidase with low enzymatic activity. FEMS Microbiol Lett, 255, 43-51.
連結：
Brevet, A., Chen, J., Leveque, F., Blanquet, S. and Plateau, P. (1995) Comparison of the enzymatic properties of the two Escherichia coli lysyl-tRNA synthetase species. J Biol Chem, 270, 14439-14444.
連結：

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