对虾白斑综合征病毒(WSSV)基因组细菌双杂交系统的构建  

Construction of White Spot Syndrome Virus(WSSV) Whole Genome Bacterial Two-hybrid System Library

作　　者：熊生良[1] 杨丰[2] 

机构地区：[1]厦门大学生命科学学院,福建厦门361005 [2]国家海洋局第三海洋研究所,福建厦门361005

出　　处：《厦门大学学报（自然科学版）》2006年第5期696-699,共4页Journal of Xiamen University：Natural Science

基　　金：国家自然科学基金(30330470);福建省自然科学基金(2003F001)资助

摘　　要：细菌双杂交系统是新近建立的一种研究蛋白质间相互作用的方法.应用细菌双杂交系统,分别以pBT、pTRG诱饵质粒构建编码WSSV基因组DNA随机片段的融合表达质粒pBT-wssv、pTRG-wssv.重组质粒共转化双杂交报告菌株XLl-Blue MRF′,通过LB-TCK平板筛选对虾白斑综合征病毒中有相互作用的蛋白质.本研究中我们构建了WSSV基因组细菌双杂交系统,用于筛选WSSV中有相互作用的蛋白,为该病毒功能基因组的研究打下良好的基础.Bacterial two-hybrid system is a newly developed method for studying protein-protein interaction. We used BacterioMatch two-hybrid system （Stratagene） to study the interaction of WSSV protein. First,the random segments （about 1 kb） of WSSV genome were inserted into the plasmids pBT and pTRG to construct recombinant plasmids pBT-wssv and pTRG-wssv respectively. Then these two plasmids were cotransformed into the competent bacteria XL1-Blue MRF＇. And a reporter gene had been embedded into this strain. Therefore,we used LB-TCK agar plate to screen the WSSV protein in order to find the interaction of them. In current research we had constructed the WSSV gene group bacterial two-hybrid system, which had been used to screen the WSSV genome to find the protein interaction. And we also built a good foundation for this viral functional genome research.

关 键 词：对虾白斑综合征病毒 细菌双杂交 pBT诱饵质粒 pTRG诱饵质粒 共转化 

分 类 号：S945.19[农业科学—水产养殖]