新成人腹泻轮状病毒J19株NSP1、NSP2和NSP3基因序列分析

Sequence analysis of the NSP1,NSP2,and NSP3 genes of a novel adult diarrhea rotavirus strain J19

目的克隆新成人腹泻轮状病毒J19株三个非结构蛋白NSP1、NSP2和NSP3基因,并分析其基因序列。方法利用一种改进的非依赖核酸序列的单引物扩增方法扩增J19株三个基因,克隆到pMD18-T载体中并进行测序。在此基础上,将J19株的NSP1、NSP2和NSP3的蛋白序列与其他轮状病毒蛋白序列进行比较分析和种系进化分析。结果J19株的NSP1、NSP2和NSP3基因为基因5、7和8,它们的全长1307个、1004个和932个核苷酸,编码395个、297个和262个氨基酸。与J19株的NSP1、NSP2和NSP3蛋白序列一致性较高的分别是B组轮状病毒KB63株(26.3%)、WH1株(46.6%)和IDIR株(29.6%)。对J19株的NSP1、NSP2和NSP3的遗传进化分析表明,J19株在进化树上的位置都靠近A、B和C组轮状病毒分支的根部,而且它比较偏向于B组轮状病毒的分支。结论J19株的NSP1、NSP2和NSP3与其他轮状病毒的相应蛋白序列存在显著差异。J19株NSP1、NSP2、NSP3的蛋白序列比较和遗传进化分析表明新成人腹泻轮状病毒与成人腹泻轮状病毒可能有共同起源;但是新成人腹泻轮状病毒与成人腹泻轮状病毒存在显著差异。

Objective To clone and sequence the non-structural proteins NSP1, NPS2, and NSP3 of a novel adult diarrhea rotavirus strain J19. Methods NSP1, NSP2, and NSP3 genes of J19 were amplified by an improved single primer sequence-independent amplification method, cloned into pMD18-T vector, and sequenced. Amino acid sequence comparisons and phylogenetic analysis of NSPI, NSP2, and NSP3 of J19 were made with the known proteins of other rotaviruses. Results Genomic segment 5,7, and 8 of J19 encoding NSP1, NSP2, and NSP3 were 1307bp,1004bp, and 932bp in length, and the deduced proteins were 395 aa,297 aa, and 262 aa respectively. NSP1, NSP2, and NSP3 of J19 showed moderately higher identities to group B rotavirus strains KB63 （26.3%）, WH1 （46.6%）, and IDIR （29.6%）, respectively. Phylogenetic analysis of NSP1, NSP2, and NSP3 of J19 showed that J19 was positioned close to the ancestral nodes of group A, B, and C lineages, although with a preferred association with group B lineages. Conclusion NSP1, NSP2, NSP3 of J19 differed significantly from those of any other known rotaviruses. Protein sequence analysis and phylogenetic analysis of NSP1, NSP2, NSP3 of J19 showed that the novel adult diarrhea rotavirus and the adult diarrhea rotavirus would originate from a common ancestral virus; but the novel adult diarrhea rotavirus differed significantly from other known adult diarrhea rotavirus.