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鳗弧菌(Vibrio anguillarum)侵染对青蛤(Cyclina sinensis)谷胱甘肽硫转移酶及其基因表达的影响
罗凯娅, 刘欣欣, 葛端阳, 潘宝平
天津师范大学生命科学学院 细胞遗传与分子调控天津市重点实验室
摘要:
在鳗弧菌胁迫下测定了青蛤血清及肝脏谷胱甘肽硫转移酶(GSTs)活力, 利用SMART cDNA文库和高通量测序方法, 筛选到青蛤σ型谷胱甘肽硫转移酶基因(CsGSTS)的全长。采用荧光定量PCR法分析CsGSTS基因的表达过程。结果表明, 青蛤的CsGSTS基因cDNA全长793bp, 编码206个氨基酸, 具典型的GST-N和GST-C结构域。血清中的GSTs活力在感染后6—24h显著升高(P<0.01), 肝脏GSTs活力在6—12h显著下降(P<0.05), 48—96h又明显上调(P<0.01); 肝脏CsGSTS基因表达水平在胁迫后3—6h降低, 24h 显著升高, 48h 达到最大值, 约为对照组的3倍。说明谷胱甘肽硫转移酶及其基因表达参与了青蛤的免疫应答反应。该研究为探索贝类的抗病害免疫机制提供了重要的实验数据。
关键词:  青蛤, 谷胱甘肽转移酶(GSTs), 酶活力, CsGSTS 基因, 基因表达
DOI:10.11693/hyhz201204008008
分类号:
基金项目:天津市科委应用基础与前沿技术重点项目资助, 12JCZDJC22800 号
EFFECT OF VIBRIO ANGUILLARUM ON ACTIVITY AND GENE EXPRESSION OF GLUTATHIONE S-TRANSFERASES IN CYCLINA SINENSIS
LUO Kai-Ya, LIU Xin-Xin, GE Duan-Yang, PAN Bao-Ping
College of Life Sciences, Tianjin Key Laboratory of Cyto-Genetical and Molecular Regulation, Tianjin Normal University
Abstract:
The activities of glutathione S-transferases (GSTs) in serum and liver were tested under Vibrio anguillarum stress. The sigma-like glutathione S-transferases of Cyclina sinensis (CsGSTS) were cloned with SMART-cDNA library and large scale EST sequencing method, then the expression of CsGSTS in liver were detected by the real-time quantitative PCR. Results showed the GSTs activities in serum were up-regulated at 6—24h (P<0.01) after infection, while in liver the activities declined at 6—12h and increased significantly at 48—96h. The full length cDNA of CsGSTS consisted of 793bp, encoding 206 amino acids with the typical GST-N and GST-C domain. The CsGSTS expression declined at 3—6h (P<0.05), and increased significantly at 24h, then reached its highest level at 48h, which was about three times higher than control group. Results showed that Vibrio may cause tissue injury at the first stage of infection, then plenty of GSTs were produced and CsGSTS expressed intensively, both participating in the immune response of the clam. This investigation provided important experimental data for the study on immunologic mechanism in C. sinensis.
Key words:  Cyclina sinensis, Glutathione S-transferases (GSTs), Enzyme activity, CsGSTS gene, Gene expression
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