蒺藜苜蓿酵母杂交cDNA文库的构建与分析  

Construction and Analysis of a Yeast cDNA Library from Medicago truncatula

作　　者：李舒文 董笛 李殷睿智 王梦迪 韩烈保[1] 张铁军 Li Shuwen;Dong Di;Li Yinruizhi;Wang Mengdi;Han Liebao;Zhang Tiejun(College of Grassland Science,Beijing Forestry University,Beijing,100083)

机构地区：[1]北京林业大学草业与草原学院,北京1000083

出　　处：《分子植物育种》2021年第23期7861-7866,共6页Molecular Plant Breeding

基　　金：国家自然科学基金面上项目(31772656)资助。

摘　　要：酵母双杂交和单杂交技术是筛选蛋白质互作或蛋白质与DNA互作的一种高效分子生物学技术,是生物大分子互作和调控的重要的手段。为了获得高容量的蒺藜苜蓿(Medicago truncatula)基因文库,为后续深入挖掘蒺藜苜蓿相关基因,改善蒺藜苜蓿品质提供基础。本研究选取了不同组织来源的蒺藜苜蓿,并对蒺藜苜蓿进行不同的激素诱导或胁迫处理,利用SMART技术成功构建高容量的蒺藜苜蓿酵母杂交cDNA文库。文库质量检测结果表明:文库滴定数为5×10^(7)CFU/m L,文库库容量为1.28×10^(7)CFU,平均插入片段长度大于1000 bp,24个克隆全部能够扩增出条带,c DNA片段重组率为100%。该文库质量较高,所包含的遗传信息完整,符合酵母杂交筛选试验要求,可应用于蒺藜苜蓿基因表达调控的研究和互作蛋白的筛选试验。Yeast two-hybrid and one-hybrid technologies are an efficient molecular biology technique for screening protein interactions or protein and DNA interactions,and are an important means for the interaction and regulation of biological macromolecules.In order to obtain a high-capacity Medicago truncatula gene library,it provides a basis for further digging the related genes of Medicago truncatula and improving the quality of Medicago truncatula.In this study,Medicago truncatula from different tissue sources was selected,and different hormone induction or stress treatments were performed on Medicago truncatula,using SMART technology to successfully construct a high-capacity Medicago truncatula yeast hybrid cDNA library.The library quality test results showed that the library titration number was 5×10^(7)CFU/mL,the library capacity was 1.28×10^(7)CFU,and the average insert fragment length was greater than 1000 bp.All 24 clones were able to amplify bands,and the cDNA fragment recombination rate was 100%.The library has high quality and complete genetic information,which meets the requirements of yeast hybrid screening test,and can be applied to the research of gene expression regulation of Medicago truncatula and the screening test of interaction protein.

关 键 词：蒺藜苜蓿 酵母杂交cDNA文库 SMART技术 不同组织来源 

分 类 号：Q943.2[生物学—植物学]