黄萎病菌诱导海岛棉酵母双杂交cDNA文库构建及评价  

Construction and Characterization of Yeast Two-Hybrid c DNA Library Derived from Roots of Gossypium barbadense Inoculated with Verticillium dahliae

作　　者：杨君[1,2] 张艳[1] 王伟巧 荣伟[1] 王省芬[1] 马峙英[1,2] 

机构地区：[1]河北农业大学农学院教育部华北作物种质资源研究与利用重点实验室河北省作物种质资源重点实验室,保定071001 [2]河北农业大学作物学博士后科研流动站,保定071001

出　　处：《生物技术通报》2014年第12期105-110,共6页Biotechnology Bulletin

基　　金：国家自然科学基金项目(31301370);国家自然科学基金项目(31301371);河北省博士后科研项目(冀人社字[2013]303号)

摘　　要：采用SMARTTM c DNA合成技术,通过同源重组方法构建了海岛棉Pima90-53经强致病力黄萎病菌诱导的酵母双杂交c DNA文库。经测定,文库容量为2.82×106,滴度为5.02×108 cfu/m L。菌落PCR显示,重组到p GADT7-Rec载体上的c DNA片段大小集中在500-1 500 bp之间,重组率93.33%。该文库完全可用于下一步互作蛋白筛选、信号转导组件确定及抗病蛋白结构和功能分析。A yeast two-hybrid cDNA library derived from roots of Gossypium barbadense cv. Pima90-53 inoculated with severe virulence Verticillium dahliae was constructed based on SMARTTM technique and homologous recombination reaction. Detection of the library indicated that its capacity and titer were 2.82× 10^6 and 5.02 × 10^8 cfu/mL, respectively. The result from colony PCR showed that the length of insert eDNA fragments ranged from 500 to 1 500 bp on average and the recombination rate was 93.33%. These results demonstrate that the library meets the demands of the standard cDNA library, which will be useful for screening interaction proteins, determining signal transduction components and analyzing the structure and function of disease-resistant proteins in the future.

关 键 词：黄萎病菌 海岛棉 酵母双杂交 CDNA文库 

分 类 号：S435.621.2[农业科学—农业昆虫与害虫防治]