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为了探究一例福建省检出的HAstV-5型星状病毒2013/Fuzhou/85毒株基因组分子结构特点,本研究采用PCR分段扩增、测序、拼接的方法,获得2013/Fuzhou/85毒株基因组序列全长6 803bp:5’端和3’端均有85bp非编码区;中间三个开放阅读框:ORF1a长2 802bp(86～2 887nt),编码非结构蛋白丝氨酸蛋白酶;ORF1b长1 548bp(2 827～4 374nt),编码非结构蛋白RNA聚合酶;ORF2长2 352bp(4 367～6 718nt),编码结构蛋白衣壳蛋白前体。目前,Gen Bank中仅有两株HAstV-5型星状病毒全基因组序列:中国辽宁毒株(JQ403108)和巴西哥亚尼亚毒株(DQ028633),2013/Fuzhou/85毒株和中国辽宁毒株核苷酸相似度最高,达94.4%。对该HAstV-5型星状病毒3个开放阅读框分别构建系统进化树,发现ORF1a与HAstV-1(JF327666)相似度最高,ORF1b和ORF2与HAstV-5(JQ403108)相似度最高,提示其有可能存在重组,用Simplot软件进行重组分析,重组位点位于2 741bp,在ORF1a和ORF1b重叠区的上游。本研究中对2013/Fuzhou/85毒株的全基因组测序和重组分析,可以为星状病毒的重组和遗传进化规律研究提供参考。
In order to explore the molecular structure of a genotype of HAstV-5 2013 / Fuzhou / 85 strain detected in Fujian Province, this study used PCR segmented amplification, sequencing and splicing methods to obtain 2013 / Fuzhou / 85 strain The full length of the genome sequence was 6 803 bp with 85 bp non-coding region at 5 ’end and 3’ end. The middle three open reading frames: ORF1a was 2 802 bp (86-2 887 nt) long and encoded a non-structural protein serine protease; ORF1b was 1 548 bp (2 827 ~ 4 374 nt), encoding a non-structural protein RNA polymerase; ORF2 was 2 352 bp long (4 367 to 6 718 nt) and encoded a structural protein capsid protein precursor. At present, there are only two GenBank HAstV-5 type astrovirus genome-wide sequences: China Liaoning strain (JQ403108) and Brazil Goiania strain (DQ028633), 2013 / Fuzhou / 85 strain and China Liaoning poisoning Strain nucleotide similarity was highest, reaching 94.4%. The ORF1a and HAstV-1 (JF327666) were found to have the highest similarity to ORF1b and ORF2 with HAstV-5 (JQ403108), suggesting that they have Recombination may exist, using recombinant software Simplot analysis, the recombination site located at 2 741bp, ORF1a and ORF1b overlap region upstream. In this study, whole genome sequencing and recombination analysis of 2013 / Fuzhou / 85 strains could provide a reference for the research of the recombinant and genetic evolution of astrovirus.