高温胁迫下匍匐翦股颖酵母cDNA文库的构建与鉴定

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1.36×107 CFU，所插入的匍匐翦股颖平均片段长度> 1 000 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。

Construction and identi?cation of a yeast c DNA library of creeping bentgrass under heat stress#br#

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 1.36×10⁷CFU, and the average length of the fragment was more than 1000bp. All the 24clones 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.