食品科学

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副溶血性弧菌整合接合元件核心基因表达对温度变化的响应

朱春花,李云霞,陈兰明*   

  1. 上海海洋大学食品学院,上海 201306
  • 出版日期:2015-07-15 发布日期:2015-07-08

Effect of Temperature on the Expression of Core Genes of Integrative and Conjugative Elements in Vibrio parahaemolyticus

ZHU Chunhua, LI Yunxia, CHEN Lanming*   

  1. College of Food Science and Technology, Shanghai Ocean University, Shanghai 201306, China
  • Online:2015-07-15 Published:2015-07-08

摘要:

采用实时荧光定量聚合酶链式反应方法分析在15~ 4 2 ℃ 温度范围内副溶血性弧菌(Vi b r i oparahaemolyticus)CHN25整合接合元件(integrative and conjugative elements,ICEs)ICEVpaChn1核心基因表达对温度变化的响应。结果表明:温度介导ICEVpaChn1元件保守模块核心基因表达发生显著变化,其中,编码进入排斥蛋白Eex(entry exclusion)的基因对温度变化最为敏感,低于或高于37 ℃的温度条件均强烈抑制eex基因表达(>10 倍)。此外,在15~37 ℃范围内,温度的升高显著激活编码整合酶Int(integrase)、接合转移蛋白TraI(transfer protein I)和TraG、DNA修复蛋白RumA基因的表达,且在37 ℃达到最大值;与其他检测基因明显不同,温度升高抑制转录抑制子SetR基因的表达,促进int等基因转录激活子SetC和SetD的积累,进而刺激切离,促进ICEVpaChn1元件的接合转移。实验结果揭示了环境温度对ICEs元件核心基因表达的影响,发现低温(<15 ℃)和高温(>37 ℃)条件均可能阻遏ICEVpaChn1元件及其携带基因信息在不同种属细菌间的接合转移。

关键词: 副溶血性弧菌, 整合接合元件, 实时荧光定量聚合酶链式反应, 基因表达

Abstract:

In this study, for the first time, we investigated the effect of temperature (15–42 ℃) on the expression of core
genes of the integrative and conjugative element (ICE, ICEVpaChn1) in Vibrio parahaemolyticus CHN25 using real time
polymerase chain reaction (real-time PCR). The results revealed distinct expression patterns of the tested genes. The eex
gene involved in the entry exclusion system was strongly inhibited by more than 10 folds when the bacterium was incubated
at the temperatures below or above 37 ℃. Along with increasing temperature in the range of 15–37 ℃, the expression of
int, traI, rumA, and traG genes was enhanced, and reached the highest level at 37 ℃ as the optimal growth temperature for
V. parahaemolyticus CHN25. In contrast, increasing temperature could significantly repress the expression of the gene
encoding a repressor protein SetR that can stimulate the activators of the int gene (SetC and SetD) and promote the conjugation
and transfer of ICEVpaChn1. In addition, self-transmissible activity of the ICE was likely inhibited when the bacterium was
grown at temperatures lower than 15 ℃ or above 37 ℃. The results of this study will facilitate better understanding of the
molecular mechanism underlying the effects of environment stresses on the conjugative transfer of ICEs.

Key words: Vibrio parahaemolyticus, integrative and conjugative elements, real-time polymerase chain reaction, gene expression

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