生物技术通报Issue(8)：108-113,6.DOI:10.13560/j.cnki.biotech.bull.1985.2015.08.016
大黄鱼Wap65-2基因成熟肽区酵母双杂交诱饵载体的构建及鉴定
Construction and Identification of a Bait Vector for the Mature Peptide ofWap65-2of Large Yellow Croaker in Yeast Two-Hybrid System
摘要
Abstract
The mature peptide ofWap65-2 gene(LcWap65-2m)in large yellow croaker(Larimichthys crocea)was used to construct the yeast two-hybrid bait vector, and its self-activation and toxic effect were measured.LcWap65-2m from the liver cDNA of large yellow croaker was amplified, and cloned into the plasmid pGBKT7. After verification by PCR amplification, enzyme digestion and sequencing, the bait vector pGBKT7-LcWap65-2m was transformed into the yeast strain Y187. The self-activation and toxic effect of the recombinant protein were detected in selective nutrition medium, and the expression of the bait protein LcWap65-2m was analyzed by Western blot.LcWap65-2m was successfully amplified and inserted into the plasmid vector pGBKT7. Western blot analysis showed that the bait protein LcWap65-2m was expressed in yeast cells;phenotypic screening tests revealed that the bait protein LcWap65-2m had no self-activation activity and toxic effect. In conclusion, the yeast two-hybrid bait vector pGBKT7-LcWap65-2m was successfully constructed, which laid the foundation for screening the protein interacted with the bait protein Wap65-2 using the yeast two-hybrid system.关键词
Wap65-2/酵母双杂交/表达/自激活/毒性Key words
Wap65-2/yeast two-hybrid/expression/self-activation/toxic effect引用本文复制引用
裘天休,李长红,陈炯..大黄鱼Wap65-2基因成熟肽区酵母双杂交诱饵载体的构建及鉴定[J].生物技术通报,2015,(8):108-113,6.基金项目
国家自然科学基金项目（31402333），高等学校博士学科点专项科研基金项目（20133305120008），宁波市自然科学基金项目（2013A610160），浙江省教育厅科研项目（理） （）
