摘要
目的应用酵母双杂交技术研究胞内氯离子通道蛋白1(CLIC1)的结合蛋白,以进一步了解其功能。方法首先将CLIC1的全长cDNA连接到pGBKT7载体中,重组质粒pGBKT7CLIC1转化入酵母菌株AH109,再将人Hela细胞的cDNA文库转化AH109细胞,用αgal检测阳性克隆。所获得阳性克隆测序后进行BLAST检索。结果用酵母双杂交方法筛选到了16个阳性克隆,其中有4个是已知蛋白的基因。结论利用酵母双杂交系统筛选到了数个结合蛋白。
Objective To investigate proteins associated with Chloride intracellular channel 1 （CLIC1） by yeast two-hybrid technique and, thus to clarify the functions of CLIC1. Methods First, the eDNA fragment of CLIC1 was inserted into the bait vector pGBKT7 in frame. The recombinant pGBKT7-CLIC1 was then transformed into yeast host strain AH109, followed with transformation of human Hela cell eDNA library. Candidate positive clones were assayed through α-galactosidase activity. Confirmed positive clone plasmids were sequenced and retrieved with Blast Program. Results 16 positive clones were obtained by yeast two-hybrid system, and 4 of them were known genes. Conclusion We obtain four CLIC1-assoeiated proteins with significant effects on human cells.
出处
《安徽医科大学学报》
CAS
北大核心
2006年第2期175-177,共3页
Acta Universitatis Medicinalis Anhui
关键词
酵母菌
双杂交系统技术
氯化物通道
yeasts
two-hybrid system techniques
chloride channels
