摘要
目的:Hcbp6是与核心蛋白相结合的未知功能蛋白基因,为了初步提示他的生物学功能和在细胞中的存在形式,采用酵母双杂交体系寻找与他相互作用的肝细胞蛋白,探讨Hcbp6的生物功能。方法:应用酵母双杂交系统3,构建Hcbp6诱饵质粒并转化酵母AH109,与含人肝细胞cDNA文库质粒的酵母Y187进行配合,在涂有x-α-gal营养缺陷型培养基(SD/Trp-Leu-His-Ade)上筛选生长。挑选蓝色克隆,提取此酵母克隆的质粒,转化大肠杆菌提取质粒DNA后进行测序,然后进行生物信息学分析。结果:筛选出4种与Hcbp6特异性相互作用的蛋白,包括Paralemmin(PALM)、Ran结合蛋白2(Ranbp2)、跨膜运输蛋白21(Tmp21)和血清白蛋白。结论:推测该蛋白可能为一分泌性蛋白,或与分泌蛋白的形成有关,进一步的生物学功能的研究需要更多的实验加以探讨。
AIM:To seek for hepatic proteins that interacted with protein encoded by Hcbp6 for exploring the biological function of Hcbp6. METHODS:Hcbp6 gene was introduced into pGBKT7, and then transformed into yeast AH109, which was mated with yeast Y187 (α type) containing liver cDNA library plasmid in 2×YPDA medium. Diploid yeast was plated on synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) containing x-α-gal. Plasmids were extracted from positive colonies, and sequence analysis was performed by bioinformatics. RESULTS:Four kind of proteins including paralemmin, Ran binding protein 2, transmembrane transporting protein and albumin were identified to interact with Hcbp6 specifically. CONCLUSION:Hcbp6 proteins may belong to or be associated with formation of secretary proteins, more study needs to be done for clarifying its biological function.
出处
《世界华人消化杂志》
CAS
2003年第4期385-388,共4页
World Chinese Journal of Digestology
