引用本文:	吕达,罗凯娅,潘宝平,高虹.青蛤(Cyclina sinensis)金属硫蛋白及硫氧还蛋白基因的克隆与表达分析.海洋与湖沼,2012,43(1):47-51.

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*青蛤(Cyclina sinensis)金属硫蛋白及硫氧还蛋白基因的克隆与表达分析*
吕达, 罗凯娅, 潘宝平, 高虹
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:

利用构建的 cDNA 文库及高通量测序方法, 获得金属硫蛋白及硫氧还蛋白基因全长。结果表明, 青蛤金属硫蛋白和硫氧还蛋白的 cDNA 全长分别为 776bp 和 804bp, 金属硫蛋白基因编码 74个氨基酸, 包含 7 个 CXC 特征结构; 硫氧还蛋白基因编码 165 氨基酸, 包含 5 个氨基酸构成的活性中心。采用实时定量 PCR 方法分析了两种基因在 Cd²⁺胁迫下的表达变化, 结果显示, 金属硫蛋白基因在 Cd²⁺胁迫下 6—12h mRNA 表达量急剧上升, 与对照组有显著性差异(P<0.01)。硫氧还蛋白基因在 Cd²⁺胁迫下 mRNA 在 6h 明显升高, 在 6—24h 与对照组出现显著性差异(P<0.05)。说明 Cd²⁺能够诱导青蛤金属硫蛋白和硫氧还蛋白基因表达的时序性升高, 二者的转录过程反映了贝类抵御重金属胁迫的分子调控过程。

关键词: 青蛤, 金属硫蛋白(MTs), 硫氧还蛋白(Trx), 基因表达

DOI：10.11693/hyhz201201007007

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, 09JCZDJC19300 号; 天津市科委应用基础研究面上项目资助, 10JCYBJC09100 号

附件

EXPRESSION OF METALLOTHIONEIN AND THIOREDOXIN GENE IN CYCLINA SINENSIS EXPOSED TO CADMIUM

Lü Da, LUO Kai-Ya, PAN Bao-Ping, GAO Hong

College of Life Sciences, Tianjin Key Laboratory of Cyto-Genetical and Molecular Regulation,Tianjin Normal University

Abstract:

The sequences of metallothionein and thioredoxin of Cyclina sinensis were cloned with the large scale EST sequencing method to built the clams C. sinensis SMART-cDNA library. Real-Time PCR (RT-PCR) technique was used to detect the expression of metallothionein and thioredoxin gene under Cadmium (Cd). The results showed that the full length cDNA of metallothionein and thioredoxin was 776bp and 804bp respectively. The open reading frame (ORF) of metallothionein encoded 74 amino acids (aa) with 7 typical sequence: CXC. The ORF of thioredoxin encoded 165 amino acids with a typical active position (Trp-Cys-Gly-Pro-Cys.) Real-Time PCR showed that the expression of metallothionein gene significant increased under Cd²⁺ stress from 6h to 12h, which was significant different from the control group (P<0.01), then it tend to normal level after 24h. The expression level of thioredoxin gene increased under Cd²⁺ intimidated at 6h. And the expression of 12h, 24h was significant different from the control group (P0.05). Our investigation showed that Cd²⁺ could induce the expression of metallothionein and thioredoxin gene, and the transcription reveals the molecular regulation mechanism in the immune defense of Cd²⁺ stress in clam of Cyclina sinensis.

Key words: Cyclina sinensis, Metallothionein (MTs), Thioredoxin (Trx), Gene expression