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目的对保存的基孔肯雅病毒WJ0601株基因组序列进行测定,并阐明该病毒与已报道毒株序列的关系。方法对基孔肯雅病毒基因组编码区分14段进行RT-PCR扩增,对非编码区采用RACE法进行扩增,将扩增产物直接进行测序,应用DNAStar软件将测序结果拼接得到基因组序列。结果与结论基孔肯雅病毒WJ0601株基因组核苷酸共11826nt,编码3722个氨基酸,其中5′端的2/3基因组编码4种非结构蛋白nsP1、nsP2、nsP3和nsP4;3′端的1/3基因组编码5种结构蛋白C、E3、E2、6K和E1蛋白。结构基因和非结构基因之间有68nt的连接区为非翻译区。病毒基因组5′端和3′端分别有76nt、526nt的非编码区。序列同源性分析结果表明,此株病毒与S27-African株的同源性最高,两者核苷酸序列同源性为99.9%,氨基酸序列同源性为99.8%,同属于(ESCA)基因型。
Objective To determine the genomic sequence of the conserved Chikungunya virus (WJ0601) strain and to elucidate the relationship between the virus and the reported strain sequence. Methods Chikungunya virus genome was divided into 14 segments for RT-PCR amplification. Non-coding region was amplified by RACE. The amplified products were directly sequenced. DNAStar software was used to splicing the genome sequence to obtain the genome sequence. RESULTS AND CONCLUSION Genomic DNA of Chikungunya virus strain WJ0601 was 11826 nt in total and encoded 3722 amino acids. The 2/3 genome at the 5 ’end encodes four non-structural proteins, nsP1, nsP2, nsP3 and nsP4. The genome encodes five structural proteins C, E3, E2, 6K and E1 proteins. 68nt of the junction between the structural and nonstructural genes is the untranslated region. The 5 ’and 3’ end of the viral genome has a non-coding region of 76 nt and 526 nt, respectively. Sequence homology analysis showed that the homology of this strain with S27-African strain was the highest, with 99.9% nucleotide sequence homology and 99.8% amino acid sequence homology (ESCA) type.