摘要
目的筛选丙型肝炎病毒非结构蛋白4B(HCVNS4B)在人白细胞cDNA文库中的相互作用蛋白。方法应用酵母双杂交技术筛选人白细胞文库中与HCVNS4B相互作用的蛋白,对筛选结果中目的蛋白的编码基因进行克隆,并构建其酵母表达载体,在酵母细胞中进行回交验证。结果共筛选出人白细胞cDNA文库中与能够与HCVNS4B相互作用的蛋白5种(溶菌酶前体、推测的人翻译起始因子、人选择素P配体、人HC7371基因和TNF-alpha转换酶)以及一个未知功能基因。对新基因成功克隆,在酵母细胞中回交验证了HCVNS4B与该基因的相互作用。结论在人白细胞cDNA文库中存在一些能够与HCVNS4B相互作用的蛋白,它们均与细胞的生长和代谢状态有着密切的关系,其具体作用机制尚有待进一步研究。
Objective To screen and identify the proteins interacting with hepatitis C virus nonstructural protein 4B （HCV NS4B） in human leukocytes cDNA library. Methods The proteins interacting with HCV NS4B were screened and identified with yeast two-hybrid in human leukocytes, and coded gene of the interest protein was cloned. After the yeast expression vector of the interest gene was constructed, back-cross was performed. Results Five colonies,including lysozyme C precursor, homo sapiens putative translation initiation factor, homo sapiens selectin P ligand, homo sapiens HC7371 gene, and TNF-alpha convertase and an unknown functional gene were screened. The unknown functional gene was cloned successfully,and the interaction between NS4B and the unknown functional gene was verified by back-cross in yeast cells. Conclusions In human leukocytes cDNA library,there are some proteins interacting with HCV NS4B. These proteins have close relationship with cellular growth and metabolism. The mechanism of the action should be further studied.
出处
《传染病信息》
2008年第3期158-160,共3页
Infectious Disease Information
