摘要
本研究利用PCR方法扩增小反刍兽疫病毒(PPRV)的衣壳蛋白(N蛋白)基因,并将其插入真核表达载体pcDNA3.1中,构建重组表达质粒pcDNA3.1-pN。利用电转的方法将该重组质粒转入非洲绿猴肾(Vero)细胞,并使用抗生素G418进行筛选。Western blot试验结果表明,N基因在Vero细胞中得到了表达,且能够随着细胞的传代稳定遗传。PPRV N基因的成功表达为小反刍兽疫的诊断、鉴定以及新型基因重组疫苗的开发奠定了基础。
In the present study,the N gene of peste des petits ruminants virus(PPRV)was amplified by PCR and inserted into the multiple clone sites of the pcDNA3.1 vector.The recombinant plasmid was transfected into Vero cells by electroporation and screened with antibiotic G418 to stably express the N protein.The results showed that the N protein was successfully expressed and stably express with the Vero cell passage as visualized in Western blot.The stable Vero cell line expressing PPRV N protein was successfully established,which would provide high quality and stable viral capsid protein for the development of diagnostic method and of recombinant vaccines.
作者
贾雪霞
刘莹
孟德梅
樊振川
JIA Xuexia;LIU Ying;MENG Demei;FAN Zhenchuan(State Key Laboratory of Food Nutrition and Safety,College of Food Science and Engineering,Tianjin University of Science&Technology,Tianjin 300457,China;Institute of Health Biotechnology,Tianjin University of Science&Technology,Tianjin 300457,China;College of Biotechnology,Tianjin University of Science&Technology,Tianjin 300457,China)
出处
《中国动物传染病学报》
CAS
北大核心
2022年第2期56-60,共5页
Chinese Journal of Animal Infectious Diseases
基金
国家重点研发计划项目资助(2017YFD0500902)。
关键词
小反刍兽疫病毒
衣壳蛋白
稳定细胞系
表达
Peste des petits ruminants virus
capsid protein
cell line
expression
