PreS-Tat真核表达载体的构建及表达

Construction and expression of eukaryotic vector encoding PreS-Tat

目的:构建pEGFP-PreS-Tat嵌合载体并在真核细胞中表达。方法:提取人乙型肝炎病毒并用PCR法扩增出PreS片段,定向克隆入pEGFP-C3载体,在PreS下游连接合成的Tat序列,构建成功的pEGFP-PreS-Tat质粒经脂质体转染Hela细胞,Westernblot鉴定目的蛋白的表达。结果:酶切和测序结果表明成功地构建了pEGFP-PreS-Tat嵌合载体,WesternBlot结果表明该载体能在Hela细胞中表达pEGFP-PreS-Tat融合蛋白。结论:成功构建pEG-FP-PreS-Tat载体并表达出相应的融合蛋白,为研究该融合蛋白的功能奠定了基础。

Aim：To construct the chimeric vector PreS-Tat and express it in the eukaryotic cells.Methods：After amplifying the HBV PreS gene,the products were cloned into the pEGFP-C3 vector.The synthetic Tat sequences were inserted into the downstream of the PreS.The resultant recombinant vectors were transfected into eukaryotic cell Hela by lipofectamine.Western blot was used to certify the purpose protein.Results：The recombinant plasmid was confirmed by restriction endonuclease and sequencing.The pEGFP-PreS-Tat fusion protein was successfully expressed in the Hela cells.Conclusion：pEGFP-PreS-Tat has been constructed and the fusion protein is expressed successfully,which lays the foundation for studying the functions of the protein.