羊痘病毒80、81基因真核表达载体构建及鉴定

Construction and identification of eukaryotic expression vectors for sheep pox virus genes 80 and 81

试验旨在探究羊痘病毒80、81基因的生物学功能。利用在线软件对80、81基因进行生物信息学分析,以此为基础通过PCR技术扩增80、81基因,与真核表达载体pCDNA3.1构建重组质粒;重组质粒转染Vero细胞,CCK-8测定重组质粒在细胞中表达蛋白的毒力。结果显示:羊痘病毒80基因蛋白以α螺旋为主,表现出亲水性,具备功能酶位点;羊痘病毒81基因蛋白以无规则卷曲为主,表现出疏水性,也具有功能酶位点。以扩增80、81基因为模板成功构建pCDNA3.1-80、pCDNA3.1-81真核表达载体,CCK-8未检测到80、81基因在细胞中表达的蛋白具有毒力。研究表明,80、81基因具有功能位点,在转染细胞中未表现出毒力。

The aim of the study was to investigate the biological function of sheep pox virus genes 80 and 81.Bioinformatics analysis of genes 80 and 81 was performed using online software.Based on the analysis,the genes 80 and 81 were amplified by PCR technology,and the recombinant plasmid was constructed with the eukaryotic expression vector pCDNA3.1.The recombinant plasmid was transfected into Vero cells,and the virulence of the protein expressed by the recombinant plasmid was measured by CCK-8.The results showed that the sheep pox virus gene 80 protein was mainlyα-helix,which showed hydrophilicity and functional enzyme sites.Sheep pox virus gene 81 protein is mainly random curled,showing hydrophobicity,and also has functional enzyme sites.pCDNA3.1-80 and pCDNA3.1-81 eukaryotic expression vectors were successfully constructed using amplified the genes 80 and 81 as templates.CCK-8 did not detect the virulence of the proteins expressed by the genes 80 and 81 in cells.The indicates that the genes 80 and 81 has functional sites and did not show virulence in transfected cells.