中国分离的甲属病毒YN87448毒株非结构区基因的核苷酸序列测定及分析  

Complete nucleotide sequence of the nonstructural gene of Alphavirus YN87448 strain isolated in China and its relationship to other Sindbis viruses

作　　者：周国林[1] 梁国栋[1] 李蕾[1] 付士红[1] 李福胜[1] 金奇[1] 张海林[2] 黄文丽[2] 侯云德[1] 

机构地区：[1]中国预防医学科学院病毒学研究所病毒基因工程国家重点实验室,北京100052 [2]云南省流行病防治研究所

出　　处：《中华实验和临床病毒学杂志》1999年第4期314-320,共7页Chinese Journal of Experimental and Clinical Virology

基　　金：国家自然科学基金!项目编号 :3 982 3 0 0 2

摘　　要：目的　研究我国首例从发热病人血清中分离的YN87448病毒的分子致病基础。方法　甲属病毒基因序列保守区设计引物,用逆转录聚合酶链反应(RTPCR)方法扩增,再将扩增片段克隆到pGEMT载体。测定YN87448毒株非结构基因序列。结果　YN87448病毒非结构区基因序列为7613个核苷酸(不包括5′端帽子结构),编码非结构蛋白1(nsP1),非结构蛋白2(nsP2),非结构蛋白3(nsP3),非结构蛋白4(nsP4);有一个起始密码子(ATG)位于第60位碱基;含有2个终止密码子(TGA),分别位于基因组nsP3基因和nsP4基因的末端。YN87448病毒非结构区基因与辛德毕斯病毒家族中唯一能致成年小鼠死亡的S.A.AR86株相应基因的核苷酸序列同源性为988%,但YN87448毒株不致成年小鼠死亡。与S.A.AR86株相比,基因结构上有三点明显差异:YN87448毒株在nsP3区位于基因组5256bp5309bp含54个核苷酸的插入;位于基因组5603bp处3个核苷酸(AGT)的缺失;在nsP3和nsP4之间有乳白突变终止密码子(TGA)。此序列已输入GeneBank,序列号为AF103734。结论　YN87448为一株新的辛德毕斯病毒株。Objective To determine the complete nucleotide sequence of the nonstructural gene of YN87448 virus stain which was firstly isolated from a female patient with fever in Yunnan Province in 1986, and identified as a member of Alphavirus by the serological method.Methods The complete nucleotide sequence of the nonstructural region gene of YN87448 virus strain was determined with nine clones, which were obtained by using reverse transcription and polymerase chain reaction (RT PCR), and by linking nine overlapping fragments into pGEM T vector respectively.Results The complete nucleotide sequence of nonstructural gene of YN87448 virus strain was 7 613 nucleotides long exclusive of the 5'cap, encoding four nonstructural proteins, nsP1, nsP2, nsP3, nsP4, and contained one initiator(ATG) and two stop codons (TGA). In comparison with the consensus sequence of S.A.AR86, the homogeneity of the nucleotide sequence between YN87448 virus strain and sindbis like virus isolate S.A.AR86 was 98.8%. YN87448 virus strain can not produce a fatal disease in adult mice. In comparison with the consensus sequence of S.A.AR86, there is a 54 nucleotide insertion from 5 256 bp to 5 309 bp, 3 nucleotide (AGT) deletion at 5 603 bp in nsP3 region, and an opal termination codon between the nsP3 and the nsP4 genes in YN87448. virus strain. This sequence has been put into Gene Bank and No. is AF103734.Conclusion YN87448 is a new sindbis like virus strain.

关 键 词：甲病毒属 分子克隆 聚合酶链反应 碱基序列 

分 类 号：R373[医药卫生—病原生物学]