基因Ⅰ型和Ⅲ型日本脑炎病毒sfRNA亚型鉴定及遗传演化分析

Subtype identification and phylogenetic analysis of sfRNA of genotypes Ⅰ and Ⅲ Japanese encephalitis virus

旨在探究基因Ⅰ型和Ⅲ型日本脑炎病毒(Japanese encephalitis virus,JEV)感染细胞产生的黄病毒亚基因组RNA(subgenomic flaviviral RNA,sfRNA)亚型与演化趋势。应用高分辨率Northern blot检测Ⅰ型和Ⅲ型JEV感染细胞产生sfRNA亚型特征;对美国国立生物技术信息中心(NCBI)收录的99条JEV sfRNA序列通过Mega X软件进行多序列比对及遗传演化分析,用Megalign软件进行核苷酸相似性比较及变异分析。结果表明:Ⅰ型和Ⅲ型JEV感染细胞产生特征相似的4种亚型sfRNA;JEV sfRNA序列10440、10673、10853位核苷酸差异具有基因型特异性(位点说明以JEV HEN0701为例,GenBank:FJ495189.1);Ⅰ型10477位核苷酸的变化相较于Ⅲ型更有利于sfRNA PK1结构的碱基配对;Ⅰ型和Ⅲ型sfRNA组间核苷酸序列相似性为79.2%~100%,Ⅰ型sfRNA组内核苷酸序列相似性为81.6%~100%,Ⅲ型sfRNA组内核苷酸序列相似性为85.6%~100%;Ⅰ型和Ⅲ型sfRNA核苷酸变异以碱基转换为主。本研究发现JEV sfRNA存在一定差异,但同种基因型sfRNA遗传距离接近,结果为揭示JEV基因型差异提供了新思路。

This research was to identify the differences between genotypesⅠandⅢJapanese encephalitis virus in sfRNA produced in host cells.The characteristics of the subtype sfRNA produced in the genotypesⅠandⅢJEV infected cells were determined using the high resolution Northern blot.99 sfRNA sequences collected from NCBI were analyzed using the MEGA X software for multi-sequence alignment and genetic evolution analysis.Nucleotide similarity comparison and mutation analysis were performed using the Megalign software.The results showed that genotypesⅠandⅢJEV-infected cells produced 4 subtypes of sfRNA.The differences in the JEV sfRNA sequences at nucleotides 10440,10673 and 10853 were genotype-specific(The locus was taken as an example by HEN0701,Genbank:FJ495189.1).The nucleotide at position 10477 in the JEV genotypeⅠgenome was more conducive to base pairing of the sfRNA PK1 structure than genotypeⅢ.The nucleotide sequence similarity between genotypeⅠand typeⅢof sfRNA was 79.2%~100%.The nucleotide sequence similarity in genotypeⅠof sfRNA was 81.6%~100%.And the nucleotide sequence similarity the genotypeⅢof sfRNA was 85.6%~100%.GenotypesⅠandⅢsfRNA nucleotide mutations were mainly base conversions.The present study revealed that there were some differences in JEV sfRNA,but the same genotype sfRNA were genetically conserved;which provided new ideas for research on the differences between JEV genotypes.