創傷弧菌（Vibrio vulnificus）是一種嗜鹽性的革蘭氏陰性菌，會引起原發性敗血症、傷口感染和急性下痢。
　　在創傷弧菌CKM-1核酸序列分析中，我們發現兩段有趣的基因(polS、polR)。這兩個基因排列形成一個operon，且序列與二元訊息傳遞系統有很高的相似度。PolS包含His與具有histidine kinase 的感應蛋白質有很高的相似度。PolR為σ54 類似轉錄活化因子，可以加強含有σ54 啟動子的基因轉錄，與regulatory蛋白質有很高的相似度。
　　我們實驗室學長在研究創傷弧菌致病機轉的過程中，由北方點墨法發現創傷弧菌會形成polS-polR bicistronic轉錄片段; 但當菌體暴露在血清時，polR會形成monocistronic轉錄片段，藉由本身的啟動子，將PolR誘發表現。因此，在本研究中，利用RT-PCR方法更加確認當菌體暴露在血清時，PolR可以被誘發表現而PolS不能。此外，再利用EMSA的方法來研究PolR是否可以自我調節本身的基因及σ54是否可以與預測的σ54 啟動子位置結合。並將PolS及PolR蛋白質表現純化，以利進一步探討PolS是否可以將PolR磷酸化。

　　Vibrio vulnificus is a marine Gram-negative bacteria that is recognized as a cause of fulminant primary septicemia, wound infections and acute self-limiting diarrhea. In our primary study, two genes (polS and polR) were cloned from vibrio vulnificus CKM-1. These two genes appeared to be arranged in an operon and shared sequence similarity with members of two-component signal transduction systems. PolS shows sequence homologous to sensor proteins and also contains a histidine residue highly conserved in the histidine kinase family of sensor protein. PolR is homologous to other regulatory proteins that bind to specific upstream activating elements to enhance transcription of genes with σ54 promoter.
　　Our previous northern blot analysis also indicated that PolR could be expressed as a bicistronic transcript from PolS; nevertheless PolR could also be markedly induced as a monocistronic transcript from its own promoter during exposure of bacteria to serum. In this study, reverse transcription polymerase chain reaction experiments were performed to confirm that PolR but not PolS could be specifically induced during exposure of bacteria to serum. In addition, whether PolR could autoregulate itself by binding its upstream region, which locates in PolS gene structure region, or σ54 could bind the expected σ54 promtor would be examined by electrophoresis mobility shift assay. Both PolS and PolR would be overexpressed and purified, and whether PolR could be phosphorylated by PolS or not would be investigated.

Amaro, C.E., Biosca, E.G., Esteve, C., Fouz, B., and Toranzo, A.E. Comparative study of phenotypic virulence properties in Vibrio vulnificus biotype I and biotype II obtain from European eel farm experiencing mortalitie. Dis Aguat. Org. 13, 29-35, 1992
Arora, S.K., Ritchings, B.W., Almira, E.C., Lory, S., and Ramphal, R. A transcriptional activaotor, FleQ, regulates Mucin adhesion and flagellar gene expression in Pseudomonas aeruginosa in a cascade manner. J. Bateriol. 179, 5574-5581, 1997
Chang, C., and Stewart, R.C. Two component system. Plant Physiol. 117, 723-731, 1998
Chuang, Y.C., Young, C.Y., and Chen, C.W. Vibrio vulnificus infection. Scand. J. Infect. Dis. 21, 721-726, 1989
Chuang, Y.C., Young, C.Y., Liu, C.Y., Lan, C.K., and Huang, A.H. Vibrio vulnificus infection in Taiwan: report of 28 cases and review of clinical manifestations and treatment. Clini. Infect. Dis. 15, 271- 276, 1992
Chuang, Y.C., Ko, W.C., Wang, S.T., Liu, J.W., Kuo, C.F., Wu, J.J., and Huang, K.Y. Minocycline and cefotaxime in the treatment of experimental murine Vibrio vulnificus infection. Antimicrobial Agents and Chemotherapy. 42, 1319-1322, 1998
Enrique, M., and Lopenzo, S. The σ54 bacterial enhancer- binding protein family: mechanism of action and phylogenetic relationship of their functional domains. J. Bacteriol.175, 6067-6074, 1993
Ewann, F., Locht, C., and Supply, P. Intracellular autoregulation of the Mycobacterium tuberculosis PrrA response regulator. Microbiology 150, 241-246, 2004
Flashner, Y., Weiss, D.S., Keener, J., and Kustu, S. Constitutive forms of the enhancer-binding protein NtrC:evidence that essential oligomerization determinants lie in the central activation domain. J. Mol. Biol. 249, 700-713, 1995
Heeswijk, W.C., Rabenberg, M., Westerhoff, H.V., and Kahn, D. The genes of the glutamine synthetase adenylylation cascade are not regulated by nitrogen in Escherichia coli. Mol. Microbiol. 9, 443-457, 1993
Heiyoung, P., Soumitra, K.S., and Masayori, I. Two-domain reconstitution of a functional protein histidine kinase. Proc. Natl. Acad. Sci. USA 95, 6728-6732, 1998
Jeffry, B.S., Ann, M.S., and James, M.M. Signal transduction in bacterial. Nature. 344, 395-400, 1990
Jin, S., Prusti, R.K., R oitsch, T., Ankenbauer. R.G., Nester, E.W. Phosphorylation of the VirG protein of Agrobacterium tumefaciens by the autophosphorylated VirA protein: Essential role in biological activity of VirG. Journal of Bacteriology.172, 4945-4950, 1990
Karl, E.K., and John, J.M. Distinct roles of an alternative sigma factor during both free-swimming and colonizing phases of the Vibrio vulnificus pathogenic cycle. Mol. Microbiol. 28, 501-520, 1998
Krovack, K., Baloda, S.B., Dumontet, S., and Mansson, I. Detection of potential virulence markers of Vibrio vulnificus strains isolated from fish in Sweden. Comp Immunol Microbiol Infect Dis. 17, 63-70, 1994
North, A.K., Klose, K.E., Stedman, K.M., and Kustu, S. Prokaryotic enhancer-binding proteins reflect Eukaryote-like modularity: the puzzle of nitrogen regulatory protein C. J.Bacteriol. 175, 4267-4273, 1993
Potter, C.A., Ward, A., Laguri, C., Williamson, M.P., Henderson, P.J., and Phillips-Jones, M.K. Expression,purification and characterization of full-length histidine kinase RegB from Rhodobacter sphaeroides. J. Mol. Biol. 320, 201-231, 2002
Ritching, B.W., Almira, E.C., Lory, S., and Ramphal, R. Cloning and phenotypic characterization of fleS and fleR,new response regulators of Pseudomonas aeruginosa which regulate motility and adhesion to mucin. Infect. Immun. 63, 4868-4876, 1995
Sasse-Dwight, S., and Gralla, J.D. Role of eukaryotic-type domains found in the prokaryotic enhancer receptor factor σ54 . Cell 62, 945-950, 1990
Wang, W., Carey, M., and Gralla, J.D. Polymerase II promoter activation: closed complex formation and ATP driven start site opening. Science 255, 450-453, 1992