Abstract:In order to screen proteins in pancreas cDNA library interacting with HBeAg protein and study the correlation between binding proteins and metabolic syndrome, the bait plasmid pGBKT7-HBeAg was transformed into yeast strain AH109 by LiAc. Meanwhile, the library was amplifed, purified, evaluated, and transformed into yeast strain Y187. Then, the transformed AH109 mated with Y187 containing the library plasmid, and diploid yeast cells were plated on nutrient deficiency medium SD/-Trp/-Leu/-His/-Ade and SD/-Trp/-Leu/-His/-Ade containing X-α-gal for selecting. The plasmids in diploid yeast cells were extracted and electrotransformed into E.coli DH5α. A few of selected e proteins were crossed back with bait plasmid and digested by BglⅡenzyme in order to verify whether they were able to interact with the library. The valid positive plasmids were sequenced and analyzed by bioinformatic methods. The final results were that thirteen proteins interacting with HBeAg were founded, which showed that HBeAg may interact with lots of proteins of pancreas cells and may be closely related with 2 type diabetes mellitus and hepatosteatosis.
王晓春 李朝品. 酵母双杂交初步筛选与HBeAg结合的胰腺细胞蛋白[J]. 中国人兽共患病学报, 2012, 28(9): 927-931.
Wang Xiao-chun Chao-Pin LI. Screening of proteins binding to HBeAg protein from human pancreas cells by yeast two-hybrid system. Chinese Journal of Zoonoses, 2012, 28(9): 927-931.