1株麻雀源坦布苏病毒的分离鉴定及全基因组序列分析

Identification and complete genomic sequence analysis of a Tembusu virus isolated from sparrows

从山东地区某发病鸭场周围的麻雀体内分离到1株坦布苏病毒,命名为SDS株,并对分离毒株进行毒力测定。应用RT-PCR技术对分离株全基因组进行扩增并测序,将获得的SDS株坦布苏病毒全基因组序列与已在Gen Bank发表的24株坦布苏病毒和6株其他黄病毒属病毒全基因组序列进行遗传进化分析。结果表明,该株坦布苏病毒的基因组全长为10 990 nt,包含94 nt的5’端非编码区、618 nt的3’端非编码区和一个开放阅读框(3 425个氨基酸),编码11种病毒蛋白;与Gen Bank已发表的坦布苏病毒的核苷酸序列同源性为98.2%~99.5%,氨基酸序列同源性为98.1%~99.4%,其中与鸭源WFZ株(KC990545.1)的核苷酸序列同源性最高为99.5%,与鹅源坦布苏病毒(duck eggdrop syndrome virus strain goose,JQ920424.1)和鸡源坦布苏病毒(CJD50,JF926699)的核苷酸序列同源性较低为98.9%。用Net NGlyc 1.0 Server在线软件对病毒蛋白潜在糖基化位点进行预测,结果表明在SDS株病毒多聚蛋白中共有13个潜在的糖基化位点,分别位于5个不同病毒蛋白中。

This study successfully isolated a Tembusu virus( TMUV) strain from sparrows in Shandong and named Tembusu virus SDS strain. The virulence of this virus was determined by Reed-Muench method. The complete genomic sequence of this virus was amplified by overlap RT-PCR to investigate its molecular structure and genetic characteristics. The results showed that full-length genome of this TMUV is 10 990 nt,containing an open reading frame,encoding a polyprotein of 3 425 amino acids in length which cleavaged into 11 viral proteins,and a 94 nt 5’ non-coding region and a 618 nt 3’ non-coding regions. The nucleotide homology of genomic sequences between SDS strain and other TMUV isolates ranged from 98. 2%to 99. 5%. The amino acid homology of genomic sequences between SDS strain and other TMUV isolates ranged form 98. 1% to 99. 4%. Net NGlyc 1. 0 Server online software was uesd to forecast the potential glycosylation sites. The result showed that the SDS has 13 potential glycosylation sites in five viral proteins.The results of this study provides an important theoretical basis for epidemiology of Tembusu virus.