罗氏沼虾幼苗体内一种极小病毒的序列测定与分析

罗氏沼虾幼苗体内一种极小病毒的序列测定与分析 Sequence Determining and Analysis of Extra Small Virus Isolated from Giant Fresh Water Prawn, Macrobrachium rosenbergii 张化俊 1 2 3 4 first-author 袁军法 1 2 3 4 曹铮 1 2 3 4 钱冬 1 2 3 4 王建民 1 2 3 4 石正丽 1 2 3 4 中国科学院武汉病毒研究所分子病毒学实验室 中国科学院武汉病毒研究所分子病毒学实验室 浙江省淡水水产研究所 浙江省淡水水产研究所 ,浙江,湖州,313001 ,浙江,湖州,313001 从患罗氏沼虾幼苗肌肉白浊病的幼虾体内分离到两种病毒颗粒,诺达病毒（MrNV）和直径只有15nm的小病毒颗粒（extrasmallvirus,XSV）,本文通过建立病毒cDNA文库获得XSV基因组的部分序列,在此基础上合成探针,用从病虾组织中提取的总PNA进行Northern杂交,结果表明XSV基因组至少包括两条RNA片段.利用不同的方法对XSV的两端序列进行克隆和测定,分别得到了872、831和662bp3个相互重叠的序列片段,这3个序列的阅读框编码一个相同的大小在17×103左右的蛋白. Two kinds of viral particles,Nodavirus（MrNV）and Extra Small Virus（XSV） which was only about 15 nm in diameter, were isolaed from Macrobrachium rosenbergii postlarvae suffering from white tail disease. In recent years, white tail disease（WTD） has been causing an important economic losses in cultured giant fresh water prawn, Macrobrachium rosenbergii. Two kinds of viral particles were isolated from diseased Postlarvae, Macrobrachium rosenbergii Nodavirus （MrNV） and Extra Small Virus（XSV） which was only about 15 nm in diameter. In this paper, we firstly cloned XSV partial sequence, then synthesized the probes based on CDNA library and performed Northern bolt to total RNA of diseased animals. Northern bolt analysis suggested that XSV genome contained at least two RNA segments. This result was confirmed by cDNA sequencing of XSV 5′-end and 3′-end using different methods. Three overlapped fragments but with different size in nucleotide （872831和662bp）were obtained. All three fragments shared the same open reading frame which encodes a 17×10 3 protein. 罗氏沼虾肌肉白浊病 极小病毒 序列测定 white tail disease extra small virus（XSV） sequencing S945.4 国家自然科学基金资助项目（30370057） 2004-06-01 2021-04-01 6