【摘要】 目的 :制备轮状病毒外壳蛋白VP4。方法 :从病人粪便中分离的毒株中提取病毒dsRNA为模板 ,经RT -PCR获得编码VP4截短蛋白基因片段cDNA(1- 12 5 3bp) ,将VP4基因片段克隆于pGEX - 4T - 2融合表达载体中 ,经dot印迹筛出阳性重组子 ,重组质粒转化大肠杆菌BL2 1(DE3)。工程菌经IPTG诱导 ,11%聚丙烯酰胺凝胶电泳。结果 :Western印迹分析表明 ,工程菌能正确表达VP4蛋白片段 ,且具有良好的抗原性。结论 :为研制轮状病毒重组疫苗、亚单组疫苗、转基因植物口服疫苗做了前期准备。更多还原

【Abstract】 Objective:Preparation of rotavirus VP4 protein.Methods:RNA of rotavirus were extracted from patients′ faeces and were transcribed into cDNA by RT-PCR.The VP4 gene fragment about 1253bps was cloned into pGEX-4T-2,and positive bacteria were selected by dot blot.VP4 protein were expressed with GST in E.coli BL21 induced by IPTG and analysised by 10% SDS-PAGE.Results:VP4 protein had fine biological activity tested by western blot.Conclusion:This study provide significant basements for rotavirus vaccine research.更多还原