汉滩病毒截短核蛋白在大肠杆菌中的表达及其初步应用

利用逆转录 聚合酶链反应 (RT PCR)从急性期HFRS病人外周血清总RNA中扩增获得汉坦病毒S片段部分编码基因 (SA) ,使用NCBIBlast软件对排分析证实为汉滩型 ,申请GenBank号 :AY42 5 61 2。将该片段克隆入原核表达载体 ,并在大肠杆菌中高效表达。所表达的蛋白分子量约为 490 0 0kDa,重组蛋白在菌体中主要以包涵体形式存在。Western blot分析表明该核蛋白具有较好的抗原活性。该蛋白在肾综合征出血热 (HFRS)的血清学诊断中具有一定的价值。

Expression of Truncated Region of S Genome Segment of Hantaan Virus in E.coli and Preliminary Use of This Recombinant Protein

Objective\ Cloning and expression of Hantaan virus S gene truncated region(SA) and using it in rapid diagnosis of HFRS.Methods Obtained SA by RT PCR from the HFRS patient peripheral blood and cloned into GST fusion protein expression vector PGEX4T 2 by PCR and restriction digest methods,the recombinant protein was examined by immunoblot with polycloned antibodies.Results The molecular weights of SA was 49000 kDa and mainly expressed as inclusion body in Escherichia coli .Western blot analyses demonstrated that this recombinant protein presented the specific antigenicity\ Conclusion The prokaryotic expression of portions of the NPs of this specific Hantaan virus could be used to generate readily an efficiency in diagnostic assays for HFRS.;