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目的通过基因重组方式获取高浓度重组诺如病毒GⅡ4型衣壳蛋白。方法将诺如病毒GII 4型衣壳蛋白基因片段插入pHT A载体中,测序鉴定正确后,将重组质粒转化到MAX Efficiency~ DH10Bac~(TM)感受态细胞,表达目的蛋白。重组蛋白用Ni-NTA His蛋白亲和柱纯化,用诺如病毒检测试剂盒进行鉴定。结果 SDSPAGE结果表明成功表达了分子量约为60 kD的重组蛋白,经诺如病毒检测试剂盒鉴定,表达产物为重组诺如病毒衣壳蛋白,并且具有较好的免疫原性。结论构建了诺如病毒GII 4型衣壳蛋白表达载体,并获得重组诺如病毒GII 4型衣壳蛋白。
Objective To obtain high concentration recombinant Norovirus GⅡ4 capsid protein by genetic recombination. Methods The Norovirus GII type 4 capsid protein gene fragment was inserted into pHT A vector. After sequencing and identification, the recombinant plasmid was transformed into MAX Efficiency ~ DH10Bac ~ (TM) competent cells to express the target protein. The recombinant protein was purified with Ni-NTA His protein affinity column and identified with norovirus detection kit. Results SDSPAGE results showed that recombinant protein with molecular weight of about 60 kD was successfully expressed and identified by norovirus detection kit. The expressed product was recombinant norovirus capsid protein with good immunogenicity. Conclusion The Norovirus GII type 4 capsid protein expression vector was constructed and the recombinant Norovirus GII type 4 capsid protein was obtained.