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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 CTB-Eg vaccine containing intramolecular adjuvant influenza virus polytope, and to study its transfection efficiency in HEK293T cells. Methods The influenza virus multi-epitope gene CTB-Eg containing intramolecular adjuvant was designed and synthesized by bioinformatics software and inserted into eukaryotic vector pEGFP-C2 to obtain recombinant plasmid pEGFP-C2 / CTB-Eg HEK293T cells were transfected by lipofectamine. The transfection efficiency was detected by fluorescence microscopy and PCR. Results The eukaryotic expression plasmid pEGFP-C2 / CTB-Eg was successfully constructed and transiently transfected in HEK293T cells. The transfection efficiency was between 50% and 70%. Conclusion The CTB-Eg fusion gene was successfully designed and constructed in this study. The transfection efficiency of CTB-Eg fusion gene in HEK293T cells was high, which laid a solid foundation for the antiviral effect of influenza virus multi-epitope nucleic acid vaccine CTB-Eg in mice.