HBV多表位复合基因在NS-1细胞的表达

田泽维; 董文其; 王萍; 吴英松; 李林海; 李明; 黄建生

HBV多表位复合基因在NS-1细胞的表达

Expression of multiepitope of HBV in NS-1 cell

摘要

摘要:
目的研究HBc颗粒为呈现载体的HBV多表位复合基因在哺乳动物细胞NS-1内的表达。
方法 PCR扩增HBc1~72aa及93~183aa编码序列并合成HBV多表位复合基因, 定向克隆至pcDNA3.1(+)。经双酶切及核苷酸序列测定等方法筛选阳性克隆。阳性质粒以Lipofectamine介导转染NS-1细胞, 通过ELISA及间接免疫荧光等方法检测细胞表达产物。
结果双酶切及测序结果证实阳性质粒正确克隆了约890bp的HBc与HBV多表位复合基因的杂合基因。重组子成功转染NS-1细胞并表达HBc-Mep融合蛋白。
结论 HBc颗粒为呈现载体的HBV多表位复合基因在哺乳动物细胞NS-1内正确表达目的蛋白。为研究pHBc-Mep作为DNA疫苗的免疫活性奠定基础。为后期检测核酸疫苗pHBcMep的细胞免疫效果提供靶细胞。

Abstract:
Objective To study the expression of chimeric gene of the multiepitope of HBV and HBc in NS-1 cells.
Methods The genes encoding HBc1-72aa and 93-183aa were amplified with PCR, and multiepitope gene was synthesized. The genes were cloned into pcDNA3.1(+). The recombinant was verified by the restrict digestion and sequencing, and then transfected into NS-1 cells with lipofectin. The combined protein was detected by ELISA and IFA.
Results The chimeric gene, about 890 bp, was obtained by the restrict digestion and verified by sequencing. The cells transfected with the recombinant expressed the combined protein successfully.
Conclusion The combined protein (HBc-Mep) was expressed successfully in NS-1 cells. It laid the foundation for studying the recombinant using in DNA immunization.

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