摘要
利用 PCR方法 ,从阴离子交换蛋白 1(AE1)全长 c DNA中扩增出约 35 0 bp c末端 c DNA片段。测序后将其克隆至 p GADT7载体上。用醋酸锂法将构建好的 p GADT7- AE1- c-末端转染酵母菌 AH10 9,观察其在选择性培养基上的表达情况。结果表明 ,获得了 35 0 bp AE1c-末端 c DNA,p GADT7- AE1- c-末端对酵母无毒性 ,不能激活检测基因 。
In this study, about 350bp cDNA fragment was amplified by PCR. After being sequenced, the AE1 c end gene fragment was cloned into EcoRⅠ PstⅠ site of pGADT7 to form AD ends in the yeast two hybrid system. The recombinant plasmid was transformed into yeast AH109,and the expression in the yeast was observed.The results demonstrate that AE1 c end was obtained. pGADT7 AE1 c end has no toxic effect on the yeast. It can serve as a target gene of yeast two hybrid system.
出处
《生物医学工程学杂志》
EI
CAS
CSCD
2002年第2期274-286,290,共14页
Journal of Biomedical Engineering
基金
国家自然科学基金资助项目 (3 9970 2 91)
