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SARS冠状病毒核蛋白与mGM-CSF 融合质粒的构建及表达

DOI: 10.3969/j.issn.1674-6309.2005.02.002

Keywords: SARS冠状病毒,N蛋白,粒细胞-巨噬细胞集落刺激因子,融合质粒

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Abstract:

目的构建SARS冠状病毒(SARS-COV)核蛋白(N)与mGM-CSF的融合真核表达质粒,借助mGM-CSF的免疫佐剂作用,提高SARSDNA疫苗的免疫效果。方法采用PCR方法扩增N和mGM-CSF基因片段,用分子克隆和基因融合的方法将目的片段定向插入真核表达载体中,构建融合表达质粒pVAX1/N-mGM-CSF,并以融合质粒转染细胞,用免疫细胞化学和WesternBlot的方法鉴定融合质粒在体外的表达。结果经酶切鉴定及DNA序列测定证实融合质粒构建正确,细胞转染实验表明,融合质粒能在COS-7细胞中表达。结论成功构建SARS-COV核蛋白与mGM-CSF的融合表达质粒,重组质粒能在真核细胞内表达。

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