镜鲤骨骼肌酵母双杂交cDNA文库的构建及鉴定  

Construction and Identification of a Cyprinus carpio Linnaeus Skeletal Muscle Yeast Two-hybrid Library

作　　者：王书[1] 张研[1] 蒋丽[1] 赵紫霞[1] 孙效文[1] 

机构地区：[1]中国水产科学研究院,北京100141

出　　处：《生物技术通报》2012年第11期127-132,共6页Biotechnology Bulletin

基　　金：中央级公益性科研院所基本科研业务费专项资金(2010C017)

摘　　要：为了研究蛋白质之间的相互作用,利用Gateway技术构建了镜鲤(Cyprinus carpio Linnaeus)骨骼肌酵母双杂交cDNA文库。经检测表明,构建的初级cDNA文库的库容量为8×106CFU;酵母双杂交cDNA文库的库容量为6.64×106CFU,插入片段集中在1-2 kb之间,具有较好的多态性。随机挑取的24个克隆没有空载体,全部发生了重组。较高的库容量、较长的插入片段以及较高的重组率保证了文库的完整性和覆盖度。镜鲤骨骼肌酵母双杂交cDNA文库的构建为克隆全长目的基因及研究影响骨骼肌发育的信号传导通路奠定了基础。A yeast two-hybrid cDNA library of Cyprinus carpio Linnaeus skeletal muscle was constructed using the CloneMinerTMcDNA Library Construction Kit.The results showed that the primary library contains 8×106 independent clones and the yeast two-hybrid library contains 6.64×106 independent clones.The sizes of most inserts were 1-2 kb in length.All randomly selected 24 clones have recombination.High storage capacity,longer insert fragment and higher recombination rate ensured the integrity and coverage of the library.Construction of this yeast two-hybrid library of Cyprinus carpio Linnaeus skeletal muscle laid the foundation for cloning of the full-length genes in Cyprinus carpio Linnaeus and studies on signal transduction pathway of skeletal muscle growth.

关 键 词：镜鲤 酵母双杂交 GATEWAY技术 CDNA文库 

分 类 号：S917.4[农业科学—水产科学]