【摘要】 目的 :初步了解重症腹泻轮状病毒株NSP4基因特征及变异情况 .方法 :在 2 0 0 2年 10 0例秋冬季腹泻患儿轮状病毒分子流行特征的研究中 ,确诊为轮状病毒腹泻的 10例患儿的粪便标本 ,用PCR方法对其NSP4基因进行测定 ,并挑选出一株重症腹泻轮状病毒毒株 (简称 0 2′K1) ,采用一步法RT -PCR对其NSP4基因进行扩增获得NSP4cDNA ,利用银染测序方法对所扩增获得NSP4cDNA进行测定 .采用Winegene2 31核酸分析软件对 0 2′K1株NSP4核苷酸序列进行翻译 ,同时利用Omiga生物软件对 0 2′K1株、ST3株 (人轮状病毒G4血清型 )及代表NSP4四个主要基因型的之一的Wa株 (简称Wa - 1株 )、Wa -a株、Wa -v株的核苷酸和氨基酸序列进行比较 ,分析其同源性及 0 2′K1株的NSP4氨基酸变异位点 .结果 :(1)经一步法RT -PCR扩增获得的NSP4cDNA约 72 5bp ,NSP4基因全长 6 86个核苷酸 ,含有一个开放读码框架 ,编码一个由175个氨基酸组成的蛋白 ;(2 ) 0 2′K1株与ST3株、Wa - 1株、Wa -a株、Wa -v株的核苷酸和氨基酸同源性均达 90 %以上 ;(3)与ST3比较 ,0 2′K1株的NSP4氨基酸序列有 14个位点发生了变异 ,即aa16、aa2 6、aa6 0、aa72、aa76、aa12 4、aa135、aa14 0、aa14 1、aa 14 5、aa14 6、aa16 1、aa16 2、aa16 9.结论 :(1) 0 2′K1株?更多还原

【Abstract】 Objective: To study the genetic characteristics and genetic variation of the nonstructural protein NSP4 of Human rotavirus strains. Methods: One human rotavirus strain (02′K1) causing serious diarrhea was obtained from the study on rotavirus molecular epidemiologic features among 100 infants with diarrhea from September to December in 2002. The nucleotide sequence of NSP4 gene was amplified by RT-PCR and was then sequenced. The homology of the nucleotide sequences of the NSP4 genes of 02′K1, ST3, Wa-1, Wa-a and Wa-v strains was analyzed. Results: (1) Fragment 725bp in length was amplified. The NSP4 gene was found to be 686 nucleotides in length, with a single open reading frame encoding a protein composed of 175 amino acids. (2) The nucleotide sequences of the NSP4 gene were highly similar among 02′K1, ST3, Wa-1, Wa-a and Wa-v strains, with a homology greater than 90%. (3) 14 unique conserved amino acids of 02′K1 differing from ST3 strains at aa16, aa26, aa60, aa72, aa76, aa124, aa135, aa140, aa141, aa 145, aa146, aa161, aa162 and aa169 were identified within NSP4. Conclusion: (1) 02′K1 fell into Wa allele. (2) There were 14 unique conserved amino acids of 02′K1 differing from ST3 strains.更多还原