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目的在Ｅ．ｃｏｌｉ中表达ＨＣＶ ＲＮＡ聚合酶，研究其可溶性的条件，进一步研究它的抗原性，为研究其活性打下基础。方法构建表达载体 ｐＱＥ－５Ｂ－ＦＬ和 ｐＱＥ－５Ｂ－△Ｃ２１。在 Ｅ．ｃｏｌｉ（Ｍ１５）菌株中表达。在不同诱导条件下分析其可溶性，在获得ｐＱＥ－５Ｂ－△Ｃ２１可溶性蛋白后，通过Ｎｉ－ＮＴＡ柱纯化。纯化后得到的目的蛋白进行ＥＬＩＳＡ和 ｗｅｓｔｅｒｎ ｂｌｏｔ分析。结果在 Ｅ．ｃｏｌｉ中表达了 ｐＱＥ－５Ｂ－ＦＬ和 ｐＱＥ－５Ｂ－△ Ｃ２１重组蛋白，二者在 ３７℃诱导条件下均以包涵体形式存在，ｐＱＥ－５Ｂ－△２１重组蛋白在１８℃诱导条件下５０％以可溶形式存在，通过ＥＬＩＳＡ和ｗｅｓｔｅｒｎｂｌｏｔ分析，表明该蛋白具有抗原性。结论在Ｅ．ｃｏｌｉ中表达的ＨＣＶ ＲＮＡ聚合酶有良好的可溶性及抗原性。
The purpose of E. The expression of HCV RNA polymerase in E. coli, to study its soluble conditions, to further study its antigenicity, lay the foundation for the study of its activity. Methods The expression vectors pQE-5B-FL and pQE-5B- △ C21 were constructed. In E. coli (M15) strain. The solubility was analyzed under different induction conditions, and after purification of the pQE-5B-ΔC21 soluble protein, it was purified by Ni-NTA column. After purification, the target protein was subjected to ELISA and western blot analysis. The result is in E. The pQE-5B-FL and pQE-5B- △ C21 recombinant proteins were expressed in E. coli, both of which were in the form of inclusion bodies when induced at 37 ℃. The expression of pQE-5B- △ 21 recombinant protein under 50 ℃, In soluble form, the protein was antigenic by ELISA and western blot analysis. Conclusions in E. HCV RNA polymerase expressed in E. coli has good solubility and antigenicity.