白细胞介素-21与乙型肝炎病毒前S2S抗原融合蛋白在293T细胞中的表达

Expression of Fusion Protein of Interleukin-21 and Hepatitis B Virus Pre-S2 and S Antigen in 293T Cells

目的:构建白细胞介素-21(interleukin-21,IL-21)和乙型肝炎病毒前S2S抗原(S2S)的融合表达质粒,并研究其在293T细胞中的表达。方法:采用PCR方法扩增IL-21和HBV前S2S基因片段,分别克隆入pcDNA3真核表达质粒,用分子克隆方法构建融合表达质粒,并以脂质体2000转染293T细胞,分别应用ELISA法和Western Blot法检测细胞上清及细胞中IL-21和HBsAg的表达水平。结果:经酶切鉴定及DNA序列证实重组质粒内插入片段序列正确,三种重组质粒分别命名为pcDNA-IL-21、pcDNA-S2S和pcDNA-IL-21-S2S,并且重组质粒能在293T细胞内表达并分泌相关蛋白。结论:成功构建IL-21和乙型肝炎病毒前S2S抗原的融合表达质粒,重组质粒能在真核细胞内表达。

Objective： To construct eukaryotic expression plasmids carried mouse interleukin-21 （IL-21） and hepatitis B virus pre-S2 and S （HBV S2S）, and to investigate the expression of the plasmids in 293T cells. Methods： IL-21 and HBV $2S were amplified by polymerase chain reaction （PCR）, and then they were cloned to pcDNA3 eukaryotic expression plasmid. Recombinant plasmid containing both IL-21 and HBV $2S were constructed by using molecular cloning. Plasmids were transfected into 293T cells by lipofectamine 2000. Expressions of IL-21 and HBsAg in supematant and cells were tested by ELISA and Western Blot, respectively. Results： The sequences of insertion elements were confirmed by enzyme digestion and DNA sequencing. Three recombinant plasmids were named as pcDNA-IL-21, pcDNA-S2S, and pcDNA-IL-21-S2S, respectively. These plasmids could be expressed and secreted to related proteins in 293T cells. Conclusions： Recombinant eukaryotic expression plasmids containing IL-21 and HBV S2S were successfully constructed and could be expressed in eukaryotic cells.