高温胁迫下匍匐翦股颖酵母cDNA文库的构建与鉴定  

Construction and identification of a yeast cDNA library of creeping bentgrass under heat stress

作　　者：刘畅 董康挺 张雅晰 李静思 边秀举[1] 李会彬[1] 王丽宏[1] 孙鑫博[1] LIU Chang;DONG Kangting;ZHANG Yaxi;LI Jingsi;BIAN Xiuju;LI Huibin;WANG Lihong;SUN Xinbo(College of Agronomy,Hebei Agricultural University,Key Laboratory of Crop Growth Regulation of Hebei Province,Baoding 071000,China)

机构地区：[1]河北农业大学农学院/河北省作物生长调控重点实验室,河北保定071000

出　　处：《河北农业大学学报》2022年第5期51-55,F0002,共6页Journal of Hebei Agricultural University

基　　金：国家自然科学基金项目(31701955);河北省自然科学基金项目(C2021204010).

摘　　要：为深入挖掘匍匐翦股颖高温胁迫相关基因,本研究以高温处理的匍匐翦股颖叶片为试验材料,用Trizol法提取其总RNA,再用SMART技术进行双链cDNA的合成及回收纯化,并与pDONR222/pGADT7-DEST载体相连接,完成匍匐翦股颖酵母cDNA文库的构建。通过试验构建了质量较好的匍匐翦股颖酵母cDNA文库,文库容量为1.36×10^(7) CFU,所插入的匍匐翦股颖平均片段长度>1000 bp,选取的24个克隆经过PCR检测全部能够显示出条带,重组率为100%。通过使用SMART技术建立酵母cDNA文库的方法,获得了质量较高的匍匐翦股颖cDNA文库,为研究匍匐翦股颖相关基因的蛋白筛选试验做准备。In order to excavate the genes relating to heat stress in creeping bentgrass,the total RNA of creeping bentgrass treated by high temperature was extracted by Trizol method.Then the double-stranded cDNA was synthesized,recovered and purified by SMART technology,after which the cDNA was connected to pDONR222/pGADT7-DEST vector to construct the cDNA library.The cDNA library of creeping bentgrass was constructed with the capacity of 1.36×10^(7)CFU,and the average length of the fragment was more than 1000bp.All the 24 clones could show bands by PCR detection,and the recombination rate was 100%.By establishing cDNA library with SMART technology,high quality cDNA library of creeping bentgrass was obtained,which made sufficient preparation for protein screening experiment of creeping bentgrass.

关 键 词：匍匐翦股颖 CDNA文库 酵母双杂交 

分 类 号：S688.4[农业科学—观赏园艺]