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Structure and function of S9 segment of grass carp reovirus Anhui strain

机译:草鱼呼肠孤病毒安徽株S9区段的结构和功能

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A highly virulent grass carp reovirus (GCRV) strain, named GCRV-AH528, was recently purified from a diseased grass carp with hemorrhage disease in Anhui, China. GCRV-AH528 S9 segment was 1320 nucleotides in length and encoded a 418 amino acid VP6 protein. BLAST search showed that the VP6 protein owned a conserved domain belonging to the reoviral σ2 family. Phylogenetic analysis of VP6 presented that GCRV-AH528 belonged to GCRV genotype II, which was more closely related to Orthoreovirus than GCRV genotype I and genotype III. Further analysis revealed that GCRV-AH528 S9 and mammalian orthoreovirus S8 might have evolved from a common ancestral precursor and have identical mechanism in virus assembly. The expression level of vp6 gene was detected by quantitative real-time PCR (qRT-PCR). Over time, the expression level of vp6 gradually increased in Ctenopharyngodon idellus kidney cells. However, the level of vp6 expression in blood sharply increased at 4–6?days, and then decreased to a low level after GCRV-AH528 challenge (P?
机译:最近从中国安徽省患有出血病的草鱼中纯化出了一种高毒力草鱼呼肠孤病毒(GCRV)株,名为GCRV-AH528。 GCRV-AH528 S9区段的长度为1320个核苷酸,编码418个氨基酸的VP6蛋白。 BLAST搜索显示VP6蛋白具有属于呼肠孤病毒σ2家族的保守结构域。 VP6的系统进化分析表明,GCRV-AH528属于GCRV基因型II,与正咽病毒的关系比GCRV基因型I和基因型III更紧密。进一步的分析表明,GCRV-AH528 S9和哺乳动物正咽病毒S8可能是从共同的祖先进化而来,并且在病毒装配中具有相同的机制。通过定量实时PCR(qRT-PCR)检测vp6基因的表达水平。随着时间的流逝,vp6的表达水平在Ctenopharyngodon idellus肾细胞中逐渐增加。但是,血液中vp6表达水平在4-6天时急剧升高,然后在GCRV-AH528攻击后降至较低水平(P <0.05)。在所有检查过的组织中都检测到了vp6基因,而在血液,肾脏和肝脏中的水平相对较高(P <0.05)。酵母双杂交(Y2H)系统用于鉴定VP6自相互作用,而在VP6-VP6中未检测到相互作用。这项研究不仅揭示了S9片段的结构和表达模式,而且还通过酵母杂交方法分析了VP6的机制。本研究为进一步的实验设计和VP6功能的研究提供有价值的信息。

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