CMV 2b和TOMV CP酵母双杂交诱饵表达载体的构建及自激活效应检测  

Construction and Identification of Bait Vector Containing CMV 2b and ToMV CP Gene in Yeast Two-hybrid System

作　　者：王子华[1] 郑银英[1] 崔百明[1] 向本春[1] 

机构地区：[1]石河子大学农业生物技术重点实验室,石河子大学生命科学院,新疆石河子832003

出　　处：《西北农业学报》2011年第4期91-95,101,共6页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：国际科技合作项目(2008DFA30560);973计划前期研究专项(2008CB117018);石河子大学高层次人才科研启动项目(RCZX200732)

摘　　要：通过RT-PCR从CMV和ToMV新疆分离物中扩增出CMV 2b和ToMVCP这2个基因,按正确方向将目标片段分别插入诱饵载pSos中,并将重组质粒导入酵母温度敏感酵母菌株cdc25H,检测其表达产物对酵母Sos恢复系统Ras信号通路的激活作用和对酵母细胞温敏特性影响。结果表明,这2个诱饵载体构建成功,对酵母菌株cdc25H无毒性和对Sos恢复系统无激活作用。为利用酵母双杂交系统研究加工番茄病毒的感染机制和病毒-寄主相互作用及防治加工番茄病毒病奠定了基础。ToMV CP and CMV 2b genes were amplified by RT-PCR from Xinjiang isolations of CMV and ToMV respectively,and then fused with pSos.After confirmation with sequence analysis,the plasmid was transformed into the yeast cell cdc25H,and its toxicity and transcriptional activation was tested by the temperature sensitive.The results showed that two bait vector was successfully constructed,and the cdc25H transformed with bait plasmids grew well on SD/（-UL）,without toxicity and could not activate the transcription of reporter gene alone in yeast two-hybrid system.Together,the experiment lay a foundation for use of the yeast hybrid system further on the mechanisms of tomatoes virus infection,host factors involved in virus-host interaction and plant virus disease control.

关 键 词：CMV TOMV 酵母双杂交系统 

分 类 号：Q78[生物学—分子生物学]