极细链格孢菌蛋白激发子Peat1在酵母双杂交系统中转录激活活性检测  

Detection for Transcriptional Activity of Alternaria Tenuissim Protein Elicitor in Yeast Two-hybrid System

作　　者：刘延锋[1] 邱德文[1] 曾洪梅[1] 杨秀芬[1] 

机构地区：[1]中国农业科学院植物保护研究所,北京100081

出　　处：《安徽农业科学》2008年第20期8500-8501,共2页Journal of Anhui Agricultural Sciences

基　　金：国家"863"计划(2006AA10A210)

摘　　要：以PCR法从pGEX-6p-1-peaT1中扩增出peaT1基因片段,电泳回收后将peaT1基因定向克隆到plexA载体中。将诱饵载体plexA-peaT1经酶切和测序鉴定后,用PEG/LiAC法转化酵母EGY48[p8op-lacZ],并进行诱饵载体转录激活活性检测。结果表明,重组质粒经EcoR I和XhoⅠ双酶切后,琼脂糖凝胶电泳检测可见2条与预期相符的条带,表明诱饵载体构建成功。β-半乳糖苷酶活性分析表明,诱饵载体plexA-peat1无转录激活活性,对酵母菌株也无毒害作用。该诱饵载体可用于酵母双杂交系统中,为下一步筛选cDNA文库奠定了基础。The peaT1 gene fragment was amplified from pGEM-6p-1-peaT1 by PCR,and recovered target gene was cloned into pLexA vector.After digestion and sequencing,the bait vector pLexA-peaT1 was transformed into yeast strain EGY48 [p8op-lacZ] by PEG/LiAC,and the transcriptional activity of bait vector was detected.The results showed that recombinant bait plasmid pLexA-PEMG1 was constructed,for the two bands of recombinant bait plasmid in agarose gel electrophoresis were expected after digesting by restriction endonuclease EcoR I and Xho I.Therefore,the recombinant bait plasmid could be used in yeast two-hybrid system to screen a cDNA library.

关 键 词：Peat1 酵母双杂交 转录激活活性 

分 类 号：Q819[生物学—生物工程]