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Screening of serum protein of cryptogenic hepatitis binding protein by phage display system
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- Author:
- No author available
- Journal Title:
- WORLD CHINESE JOURNAL OF DIGESTOLOGY
- Issue:
- 36
- DOI:
- 10.3969/j.issn.1009-3079.2006.36.014
- Key Word:
- 隐源性肝炎;血清蛋白;结合蛋白;丙型肝炎病毒;噬菌体表面展示技术
Abstract： 目的:通过筛选隐源性肝炎血清蛋白结合蛋白,为隐源性肝炎的发病机制研究探索新途径.方法:应用噬菌体表面展示技术,将隐源性肝炎患者血清作为固相筛选蛋白,用噬菌体正常人肝细胞T7 cDNA文库进行5轮生物筛选,经噬斑PCR,对筛选克隆进行DNA序列分析,将推断的氨基酸序列在蛋白质库中进行搜索,自GenBank中获得结合蛋白的cDNA和基因组的全长序列,然后再探讨该蛋白在隐源性肝炎发病机制中的作用.结果:噬菌体经过5轮生物富集后,从随机筛选的60个克隆中得到24个阳性克隆,经序列测定后进行同源搜索,初步确定人类HCV NS5A转化调节蛋白13(NS5ATPl3)为人体肝脏中固有的与隐源性肝炎血清蛋白结合的蛋白.结论:应用T7 cDNA噬菌体表面展示技术,我们发现隐源性肝炎血清蛋白与人类肝细胞HCV NS5A转化调节蛋白13相互作用,在隐源性肝炎发生及抗病毒治疗有重要意义.
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