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目的对水痘-带状疱疹病毒(varicella-zoster virus,VZV)Oka株gC基因进行克隆及序列分析。方法取第40代Oka VZV疫苗株,反复冻融3次后,收集上清,提取病毒DNA,根据NCBI登录的Dumas株VZV序列,设计引物,扩增gC基因,与pMD19-T Vector连接后,热转化至E.coli Competent Cells JM109中,挑选阳性菌落进行测序;将得到的序列与Gen Bank中登录的其他12株VZV毒株的gC基因序列,进行核苷酸序列和氨基酸水平分析。结果 13株VZV-gC基因开放阅读框(ORF)的核苷酸长度为1 557~1 776 bp,编码的蛋白由519~592个氨基酸组成。核苷酸序列同源性在94.8%~100%之间,氨基酸序列同源性在93.9%~100%之间。R2重复拷贝数为7,Dumas、SVETA和Ellen重复拷贝数为6,LAX1为4。结论 VZV Oka株gC蛋白具有高度的保守性,与其功能的发挥密切相关。
Objective To clone and sequence the gC gene of varicella-zoster virus (VZV) Oka strain. Methods The 40-generation Oka VZV vaccine strain was obtained and repeatedly frozen and thawed three times. The supernatant was harvested and the virus DNA was extracted. According to the VZV sequence of Dumas strain NCBI, primers were designed and gC gene was amplified. After ligated with pMD19-T Vector, The positive clones were selected and sequenced. The obtained sequences were compared with the gC gene sequences of other 12 VZV strains registered in Gen Bank to analyze the nucleotide sequence and amino acid level. Results The nucleotide length of 13 open reading frames (ORFs) of 13 VZV-gC genes was 1 557-1 776 bp, and the encoded protein consisted of 519-592 amino acids. Nucleotide sequence homology was 94.8% ~ 100%, amino acid sequence homology was 93.9% ~ 100%. R2 repeat copy number is 7, Dumas, SVETA and Ellen repeat copy number is 6, LAX1 is 4. Conclusion The gC protein of VZV Oka strain is highly conserved and closely related to its function.