摘要
目的为进一步研究HCV影响糖、脂代谢机制奠定研究基础。方法扩增人胰腺cDNA文库并经纯化鉴定后，将文库质粒转化酵母菌株Y187。诱饵质粒pGBKT7-core转化酵母菌株AH109，在色氨酸缺陷型培养基（SD／Trp）上筛选阳性菌落。应用酵母双杂交系统3将阳性重组AH109菌株与重组酵母菌株Y187进行配合，在四缺培养基（SD／Trp／Leu／-His／-Ade）和铺有Xα-gal的四缺培养基上进行筛选，提取蓝色酵母菌落质粒，电转化大肠埃希菌DH5 α后再提取质粒测序，测序结果进行序列比对。结果筛选出11种与HCV核心蛋白相结合的蛋白基因，包括胰凝乳蛋白酶原B1前体、羧肽酶A1，胰蛋白酶原2、胰凝乳蛋白C、胰蛋白酶1、羧肽酶B1、驱动蛋j白家族3B、胰蛋白酶2、线粒体蛋白基因、弹性蛋白酶3A、辅脂肪酶。结论在筛选出的HCV核心蛋白结合的人胰腺细胞蛋白基因中，部分与糖、脂代谢密切相关。
Objective To identify HCV core protein binding proteins. Methods The library was amplified, purified, and then were transformed into yeast strain Y187. The reconstructed plasmid pGBKT7-core was transformed into yeast strain AH109 and screened on the nutrient deficiency medium SD/-Trp. The transformed AH 109 mated with Y 187 containing the library plasmid. The 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 DH5 α. The plasmids in DH5 α were extracted and sequenced. Results Eleven proteins, including chymotrypsinogen B 1 precursor, carboxypeptidase A1, trypsinogen 2, chymotryptic peptide C, trypsin 1, earboxypeptidase B 1, kinesin superfamily proteins 3B, trypsin 2, mitochondria protein gene, elastase 3A and colipase were found to be able to bind to HCV core protein. Conclusions Proteins related with metabolism of glucose and lipid may bind to HCV core protein.
出处
《中华肝脏病杂志》
CAS
CSCD
北大核心
2009年第7期501-504,共4页
Chinese Journal of Hepatology
基金
国家自然科学基金（30600524）
