摘要
目的 :应用酵母双杂交技术及生物信息学 (bioinformatics)技术筛选并克隆NS5A TP5蛋白结合蛋白基因1(NBP1)。方法 :应用酵母双杂交系统 3 ,构建NS5A TP5诱饵质粒 ,转化酵母AH10 9,与含人肝细胞cDNA文库质粒的酵母Y 187进行配合 ,于涂有x α gal营养缺陷型培养基 (SD/Trp Leu His Ade)上筛选生长。 结果 :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 ,新基因的开放读码框架长度为 2 40个核苷酸 (nt) ,编码产物由 79个氨基酸残基 (aa)组成 ,命名为NBP1,GenBank注册号为AY 45 92 91。结论 :NBP1的筛选与克隆 ,可为进一步研究HCVNS5A
Objective:To screen and clone the human gene of protein 1 binding with NS5A-TP5 protein (NBP1) using yeast two-hybrid and bioinformatics techniques.Methods:Yeast two-hybrid system was employed to screen a human liver cDNA library with NS5A-TP5 sequence as bait, and one unknown gene was obtatined. And was cloned by bioinformatics methods, named as NBP1.Results:The full-length DNA sequence for the NBP1 gene was of 240 nucleotides. The sequence for the NBP1 gene was deposited into GenBank, and the accession number is AY459291. Conclusion: These results will pave the way for the study of the molecular mechanism of th effects of NS5A protein of HCV and the development of new therapy for chronic hepatitis C.
出处
《中西医结合肝病杂志》
CAS
2004年第5期275-277,共3页
Chinese Journal of Integrated Traditional and Western Medicine on Liver Diseases
基金
国家自然科学基金攻关项目(No.C03011402
No.C30 0 70 689)
军队"九五"科技攻关项目(No.98D063)
军队回国留学人员启动基金项目(No.98H038)
军队"十五"科技攻关青年基金项目(No.01Q138)
军队"十五"科技攻关面上项目(No.01MB135)
