家蚕谷光甘肽-S-转移酶GSTe3基因的鉴定及其真核表达  

Identification and Eukaryotic Expression of GSTe-3 Gene of Silkworm (Bombyx mori)

作　　者：潘敏慧[1] 许湲 余泉友[1] 刘佳[1] 刘迪[1] 赵丹红[1] 鲁成[1] 

机构地区：[1]西南大学生物技术学院,农业部蚕桑学重点开放实验室,重庆400716

出　　处：《中国农业科学》2010年第4期873-880,共8页Scientia Agricultura Sinica

基　　金：国家重点基础研究计划(2005CB121000);国家高技术研究计划(2006AA10A118);国家自然科学基金项目(30771630)

摘　　要：【目的】谷胱甘肽-S-转移酶(Glutathiones S-tansferases,GSTs)是一类由多基因编码的多功能同工酶超家族,在解毒内源和外源性有毒物质中起着重要作用,家蚕GSTs的研究,可为解析家蚕解毒机制奠定基础。【方法】利用生物信息学和分子生物学方法克隆和分析家蚕GSTe3(BmGSTe3),利用RT-PCR分析该基因在家蚕体内的表达情况,利用重组杆状病毒真核表达系统在sf9细胞中真核表达BmGSTe3。【结果】在家蚕中克隆了家蚕的GSTe3,该基因由5个外显子与4个内含子组成,外显子总长度为512bp,属于昆虫特异的Epsilon家族。BmGSTe3包含N-末端和C-末端2个结构域,N-末端由β-α-β-α-β-β-α共7个结构基序组成,而C-末端则由5个α螺旋构成,在启动子上游2500bp区域内共发现了15个可能的转录调控元件。BmGSTe3的表达具有较高的组织特异性,它只在家蚕血液和头部表达。BmGSTe3在sf9细胞中表达的BmGSTe3蛋白具有较好的GSTs酶活性。【结论】BmGSTe3属于昆虫特异的Epsilon家族,其蛋白具有较好的GSTs酶活性。[Objective] Glutathione S-transferases （GSTs） are a superfamily of multifunctional enzymes found in almost all living organisms. They play important roles in detoxification of endogenous and exogenous toxic substance. It will lay a foundation for the detoxification mechanism of silkworm to study on GSTs of Bombyx mori. [Method] BmGSTe3 gene was cloned and analyzed by bioinformation and molecular biology method. The expression of BmGSTe3 was analyzed in silkworm by RT-PCR. BmGSTe3 gene was eukaryotic expressed in sf9 cells by Bac-to-Bac Baculovirus Expression System. [Result] The BmGSTe3 is composed of 5 exons and 4 introns and its CDS length is 512 bp. BmGSTe3 contains 15 possible transcription regulatory elements in the upstream of 2 500 bp and two domains, which N-terminal domain is composed of 7 motifs, namely β-α-β-α-β-β-α, while C-terminal domain is composed of 5 α helixs. BmGSTe3 has a conservative Ser catalyse site, belongs to Epsilon family members and express only in hemolymph and brain. BmGSTe3 protein has good GSTs enzyme activity. [Conclusion] BmGSTe3 belongs to Epsilon family members and its protein has good GSTs enzyme activity

关 键 词：家蚕 Bm6STe3 真核表达 GSTs酶活 

分 类 号：S881[农业科学—特种经济动物饲养]