摘要
目的设计并构建含分子内佐剂的流感病毒多表位核酸疫苗CTB-Eg,初步研究其在HEK293T细胞中的转染效率。方法通过生物信息学软件设计并合成了含分子内佐剂的流感病毒多表位基因CTB-Eg,将其插入真核载体pEGFP-C2中,获得了重组质粒pEGFP-C2/CTB-Eg;用脂质体法转染HEK293T细胞,通过荧光显微镜观察和PCR法检测其转染效率。结果成功构建了真核表达质粒pEGFP-C2/CTB-Eg,且该重组质粒能在HEK293T细胞中瞬时转染,转染效率在50%～70%之间。结论本研究成功设计、构建了CTB-Eg融合基因,其在HEK293T细胞中转染效率较高,为流感病毒多表位核酸疫苗CTB-Eg在小鼠体内的抗病毒作用研究奠定了坚实基础。
Objective To design and construct the eukaryotic expression vector of CTB-Eg fusion gene of influenza virus,and study the transfection efficiency of pEGFP-C2/CTB-Eg in HEK293 Tcells.Methods CTB-Eg fusion gene containing CTB（cholera toxin B subunit）as the molecule adjuvant was designed and synthesized.The eukaryotic expression vector of pEGFP-C2/CTB-Eg was constructed.The certified recombinant plasmid was transfected in HEK293 Tcells by Liposomal Transfection Reagent.The transfection efficiency of CTB-Eg gene was detected by fluorescence microscope and PCR.Results Expression vector pEGFP-C2/CTB-Eg was constructed successfully.The recombinant plasmid could be transfected in HEK293 Tcells and its transfection efficiency could reach to 50%-70%.Conclusion The eukaryotic expression vector of pEGFP-C2/CTB-Eg was constructed successfully and transfected in HEK293 Tcells.
出处
《西部医学》
2015年第2期182-185,189,共5页
Medical Journal of West China
基金
四川省科技厅支撑计划项目(2014SZ0036)
