創傷弧菌是屬於伺機性的病原體，具有高度侵害性的能力，因此受感染的人會有很高的死亡率。為解決此病菌感染問題，必須要進一步了解此病菌的致病原因。在大部分的細菌中，它們能夠利用雙分子調控系統偵測環境中的訊號，並且調節基因的表現。目前已經證實某些伺機性的病原菌的雙分子調控系統能夠感測活體內的環境，若抑制這些特別的雙分子調控系統，會顯著降低致病菌的致病能力。以知具有超過四十組的雙分子調控系統在創傷弧菌中，然而，這些雙分子調控系統在功能上所扮演的角色卻很少被探討。為了解決這個問題，我們設計了寡核酸微陣列基因晶片來研究在不同的環境刺激下，創傷弧菌的雙分子調控系統之基因表現情形。當創傷弧菌生長在含有巴拉刈 (paraquat)的溶液中，有許多調控子發現被活化，如gltR, phoB, uphA, pleD 和lytR分別被活化了10.6，31.8，3.2，12.3和 9.8倍，因此我選擇lytR做更進一步的研究目標。利用同源序列互換取代法來製造lytR突變株。並以聚合酶連鎖反應及南方墨點法加以確認突變株，更將突變株做功能性的分析研究。結果發現突變株在生長速度、菌落大小、游泳速度、對各種抗生素的敏感力以及對環境改變的適應力都和野生株相同。不過，此lytR突變株在含有0.05% Triton X-100的環境中，比野生株具有更快的菌體分解速度，此外，lytR突變株也展現出降低盤尼西林的毒殺效應的能力，並可減低生物膜的生成。更進一步的，我們利用即時聚合酶連鎖反應，發現LytSR在過氧化氫的環境下具有調控lrgA的能力。

Vibrio vulnificus is a pathogen with a high invasive capability that causes high mortality in infected individuals. In order to solve the problem of the bacterial infection, it is essential to understand its pathogenesis. In bacteria, the detection of environmental signals and thereafter modulation of gene expression are commonly controlled by the two-component regulatory systems (2CS). Certain 2CSs have been shown that they can sense the in vivo environments for pathogenic bacteria and thus the inactivation of them results in a significant reduction in their virulence. In V. vulnificus, there are more than 40 sets of 2CS, but the functional roles of these 2CSs are poorly defined. To address this question, we set up an oligonucleotide array to investigate the expression profiles of V. vulnificus 2CS genes under different environmental stimuli. When V. vulnificus is grown in the presence of superoxide generator paraquat, expression of several response regulator genes such as gltR, phoB, citB, pleD, and lytR were found to be activated 10.6, 31.8, 3.2, 12.3, and 9.8 folds, respectively. Therefore, we chose lytR as a model for further investigation by generating a deletion mutant using the allelic exchange technique. The lytR mutant strain was further confirmed by PCR and Southern blot analysis. Functional analysis showed that the mutant exhibits normal growth rates, colony size, swarming activity, susceptibility to various antimicrobial agents, and adaptive response to environmental changes. The discernible difference is that the lytR mutant displays a faster rate of cell lysis in the presence of 0.05% Triton X-100 comparing to the parental strain. The lytR mutant also shows a decrease in susceptibility to the killing effects of penicillin and a decline in biofilm formation. Furthermore, the use of real time PCR revealed that lrgA is regulated by LytSR under the stress of H2O2.

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