构建SARS病毒E2蛋白保护性抗原原核表达载体

Construction and expression of recombinant expression vector of SARS virus E2 protein

目的 构建SARS病毒E2蛋白保护性抗原片段的原核表达载体 ,为SARS的诊断和预防奠定基础。方法 采用人工合成法合成SARS病毒E2蛋白 3’端cDNA ;采用常规分子克隆方法将此cDNA片段克隆入原核表达载体 pBV2 2 0中 ,经温度诱导和SDS -PAGE电泳证明外源蛋白表达。 结果 合成的E2蛋白cDNA经序列测定表明 ,与报道的SARS病毒相应序列一致 ,内切酶酶切及PCR均得到 4 5 6bp的cDNA片段 ,与实验设计一致 ,证明该cDNA片段插入了 pBV2 2 0载体中 ;SDS -PAGE电泳表明有外源蛋白表达。

Objective To construct expression vector and expression in E.coli of SARS virus E2 protein.Methods The E2 gene was inserted into pBV220 vector,the recombinant vector of E2 was identificated by restriction endouclease digestion and PCR.E2 protein was expressed in E.coli via induction in 42?℃.Results The analysis for recombinant vector digested by EcoR I and Pst I and PCR demonstrated that the E2 gene was inserted into pBV220.SDS-PAGE showed that the relative molecular mass(Mr)of the expressed E2 protein was about 16600.Conclusion Expression vector of E2 protein was constructed successfully and expressed effectively in E.coli.These results provided an essential preparation for obtaining a larger quantity of recombinant E2 protein for further study.