目的获得中国流行株GII-4型诺如病毒(NoV)ORF2基因序列,为进一步研究诊断、疫苗提供基础依据。方法从GII-4型NoV粪便中提取病毒RNA,经逆转录后PCR并克隆到simple PMD18-T载体获得完整开放读码柜架(ORF2)阅读框序列。结果获得了正确序列的基因II-4,亚型(genogroup II-4,GII-4)NOV ORF2基因序列,经软件分析,3株NOV ORF2序列编码的氨基酸大小约为54KD,与VA387(GII-4)氨基酸同源性为92%-93%。ORF2编码的氨基酸序列存在3个相对保守序列。结论获得GII-4型NoV ORF2基因序列,为进一步表达病毒衣壳蛋白及Nov感染的诊断试剂及防治疫苗药物研制提供基础依据。
Objective To obtain the open reading frame 2 (ORF2) sequence of norovirus GII-4 strain for further study on diagnosis and vaccine of norovirus. Methods RNA was extracted from fecal sample. After transcription and PCR, the ORF2 gene was cloned into simple T-vector. The sequences were analyzed with DNA Star software and compared with prototype sequences retrieved from the NCBI database by Clustal X and Treeview software. Results The exact whole ORF2 gene of norovirus GII-4 strain was obtained which coding 54 KD amino acid. Three strains in this study showed 92% -93% homology with VA387 strains and 3 conserved regions. Conclusion ORF2 gene of norovirus GII-4 strain was obtained, which will lay the basis for further study on diagnosis and vaccine of norovirus.