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目的 :构建SFTSV的核蛋白核酸疫苗并探讨其免疫原性。方法:采用PCR方法扩增SFTSV核蛋白基因,并用引物引入限制性内切酶酶切位点,克隆入载体p JW4303。经酶切和测序鉴定正确的质粒,用PEI瞬时转染HEK293T细胞,用免疫印迹法鉴定核蛋白的表达。同时以质粒p JW4303作为阴性对照,免疫BALB/c小鼠,酶联免疫法验证其免疫原性。结果:成功构建质粒p JW4303-N,经Western blot证实SFTSV核蛋白能够在HEK293T细胞中表达。ELISA检测免疫后小鼠血清特异性Ig G抗体及其亚型发现,Ig G抗体及其亚型较免疫前明显升高,亚型中以Ig G2a升高为主。结论:SFTSV核蛋白核酸疫苗p JW4303-N具有良好的免疫原性,为进一步研究SFTSV核蛋白奠定了基础。
Objective: To construct nucleic acid vaccine of SFTSV and explore its immunogenicity. Methods: The gene of SFTSV nucleoprotein was amplified by PCR and inserted into restriction endonuclease site with primers to construct pJW4303. The correct plasmids were identified by restriction enzyme digestion and sequencing. HEK293T cells were transiently transfected with PEI and the expression of nucleoprotein was identified by Western blotting. At the same time, plasmid p JW4303 was used as a negative control to immunize BALB / c mice and the immunogenicity was verified by enzyme-linked immunosorbent assay. Results: Plasmid pJW4303-N was successfully constructed and the expression of SFTSV nucleoprotein was confirmed by Western blot in HEK293T cells. The results of ELISA showed that Ig G antibody and its subtypes were significantly higher than those before immunization, and Ig G2a was mainly found in the subtype. CONCLUSION: The SFTSV nuclear protein nucleic acid vaccine pJW4303-N has good immunogenicity, which lays the foundation for further study of SFTSV nuclear protein.