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目的克隆表达SARS冠状病毒的主要结构蛋白(S蛋白)。方法合成SARS冠状病毒S蛋白特异性基因片断并克隆入pET32a原核表达载体,转化BL21菌,经IPTG诱导高效表达得到重组S蛋白,并通过W estern印迹对重组蛋白质进行鉴定。结果重组蛋白质经镍柱亲和层析得到了部分纯化,免疫动物后得到SARS病毒的多克隆抗体。结论经E lisa检测,表达的重组S蛋白基本具备检测病人血清中抗SARS病毒IgG和IgM的能力,可进一步用于S蛋白功能研究与SARS诊断试剂盒的研制。
Abstract：Aim To clone and express the Spike protein of SARS-CoV.Methods In this work,clone a special gene fragment of S protein of SARS-CoV into vector pET32a.lnduce the expression efficiently by IPTG after the express vector transforms E.coli BL21.Identify the recombinant protein of gain by Western blot.Results The recombinant protein was run through a pillar of nickel to turn pure,to immunize rabbit to get the polyclonal antibody.Conclusion Examined by Elisa,the purified protein can examine the IgG and the IgM in SARS-patient′s serum.It can be further used for the study on Sprotein′s function research and the development of diagnostic method of SARS.
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基本信息:
DOI：10.16152/j.cnki.xdxbzr.2006.05.022
中图分类号:Q78;Q939.4
引用信息:
[1]颜桦,陈功,蒙世杰,等.重组SARS冠状病毒S蛋白的原核表达研究[J].西北大学学报(自然科学版),2006(05):777-780.DOI:10.16152/j.cnki.xdxbzr.2006.05.022 .
基金信息:
国家863计划SARS快速反应行动计划资助项目(2003AA2Z2102)
2006-10-25
2006-10-25