中文摘要：

构建cDNA文库是利用酵母双杂交技术开展特定cDNA分离的重要基础工作。利用Gateway技术,在成功提取辣椒总RNA并分离mRNA的基础上,合成了3种读码框的双链cDNA,通过BP反应构建其相应的入门文库,再通过LR反应将cDNA迅速且定向地重组到酵母双杂分析用目的载体pDESTTM22上,构建成高质量目的文库。经检测入门文库和目的文库滴度均达到（4.0～5.0）×10^6cfu/mL,文库总容量为（2.4～3.0）×10^7cfu,平均插入片段大小为1250bp,其质量完全满足于进一步利用酵母双杂体系分析目的基因互作蛋白cDNA。

英文摘要：

It’s an important prerequisite work to construct a cDNA library for isolating a specific cDNA using yeast two-hybrid technology.In this study,after successfully extracting total RNA and isolating mRNA of pepper,the authors synthesized three-frame cDNAs,performed BP reactions to generate corresponding entry libraries and then performed LR reactions to transfer cDNA into destination vector of pDESTTM22 directionally and quickly to form a high-quality destination library using Gateway technology.It has been detected that both entry library and destination library in this report have a high titer of （4.0 ～ 5.0） × 106 cfu/mL and contain a total clones of （2.4 ～ 3.0） × 107 cfu,with an average insert size of about 1 250 bp,it can be suitable for analyzing the interaction protein cDNA of the interested destination gene of pepper by two-hybrid system.