浙江省基因Ⅰ型乙脑病毒全基因序列的测定及分析

目的 为了获得浙江省乙脑病毒基因组详尽的资料,研究基因Ⅰ型乙脑病毒的分子特征及变异程度,为乙脑病毒分子流行病学及其基因研究提供科学依据.方法 设计特异性引物、RT-PCR分段扩增XJ69和XJP613株全基因,PER产物纯化后克隆于T载体并进行序列测定.通过生物学软件进行核苷酸序列和氨基酸序列分析和病毒的系统进化分析.结果 新分离乙脑病毒XJ69和NJP613株全基因组全长均为10 964个核苷酸,含有一个开放阅读框架,编码3432个氨基酸.与GenBank中选择的32株乙脑病毒全基因序列比较发现,其核苷酸同源为83.5%～99.2%,氨基酸总体同源性为97.5%～99.7%.通过PrM/C区段、E区段及全基因序列进行系统进化分析均显示该毒株属于基因Ⅰ型乙脑病毒.结论 新分离的乙脑病毒XJ69和XJP613株属于基因Ⅰ型,与上海三带喙库蚊分离株SH17M-07关系最为接近.

Complete genome sequence analysis of Japanese encephalitis virus newly isolated In China

Objective To study the complete genome sequence of Japanese encephalitis virus(JEV) strain X J69 isolated in ZheJiang province and explore its evolution. Methods Overlapping primers were designed according to the fuU-length genomes from C, enBank. RT-PCR was used to amplify the fragments and RT-PCR products were cloned T vector, sequenced and analyzed. Results The genome of strain XJ69 and XJP613 were 10 964 nucleotides in length with a single open reading frame encoding 3432 amino acids. Comparison of the complete geoome sequences of different JEV isolates showed XJ69 and XJP613 were 83.5%-99.2% and 83.4%-99.4% nucleotide sequence homology among them respectively, which resu]ted in 94.8%-99.7% amino acid sequence homology. Phylogetic analysis through PrM/C ,E and full-length genome showed that the X J69 and XJP613 strain belonged to ganotype Ⅰ. Conclusion The nucleoiede sequence and deduced amino acid sequence of X J69 and XJP613 strain were similar to that of those of genotype Ⅰ of Japanese encephalitis virus. It belonged to genotype Ⅰ and were close to the isolates SH17M-07.