摘要
目的对鼠诺如病毒分离株BJ102062进行全基因组序列测定，并对测序结果进行亲缘关系等分析。方法参照美国国立生物技术信息中心（TheNationalCenterforBiotechnologyInformation，NCBI）发表的MNVl（AY228235）全基因组序列设计引物，根据测定的结果设计下一步引物，分段扩增测序，最后将所测各段拼接；对分离株的全基因组序列进行分析，与人源、牛源和其他鼠诺如病毒的代表株分别进行核苷酸和氨基酸序列的比较并进行亲缘关系分析。结果鼠诺如病毒分离株BJ10-2062基因组全长7381bp，G＋C含量56．63％，A＋U含量43．33％，含3个开放读码框；全基因组序列经NCBIBI．AST比对，与美国的CR4（EU004674）分离株同源性最高，为92．7％；与其他鼠诺如病毒分离株的同源性大于88％，而与人源和牛源诺如病毒分离株的同源性均小于50％；ORF1～3编码的蛋白相对分子质量分别为187×10^3、58×10^3和22×10^3，含量最高的氨基酸分别是丙氨酸（Ala）、亮氨酸（Leu）、甘氨酸（Oly）和谷氨酰胺（Gln），BJ102062分离株各编码蛋白的氨基酸序列与其他鼠诺如病毒问的同源性明显高于人源和牛源诺如病毒；与原始分离株MNVl相比，BJ10-2062VP1衣壳蛋白第296位由赖氨酸突变为谷氨酸，导致其毒力减弱。基于全基因组核酸序列构建的亲缘关系树表明，BJ10-2062与其他鼠诺如病毒分离株均属于genogroupV，与美国的CR4和NIH-A114株亲缘关系最近，并与美国的其他NIH系列毒株一起形成一个独立的分支。结论对鼠诺如病毒BJ10-2062分离株全基因组的测序分析，为进一步研究鼠诺如病毒特性的分子生物学机制奠定了基础。
Objective To sequence and analyze the complete genome of murine norovirus strain BJ10 2062 iso- lated from Beijing, China. Methods PCR primers were designed according to the reference sequence of strain MNV1 （AY228235 quencing fragments subsequent primers finally assembled for were further designed based on the the analysis of the complete genome sequencing results. All of the BJ10-2062 strain the comparisons with representative strains of human, bovine noroviruses and other murine norovirus es. Results The genome of the murine norovirus strain BJ10-2062 comprises of 7 381 nucleotides with 56. 63% GC and 43.33% AU. The genome contains three open reading frames （ORFs） and shared 92.7% homoloty with the reference strain CR4 （EU004674, Amecica）, greater than 88%homology with other murine noro- virus strains but less than 50%homoloty with human and bovine noroviruses. The predicted proteins encoded by ORF1, 2, 3 are 187×10^3 , 58×10^3 and 22×10^3 , respectively. Amino acids Ala, Leu, Gly and Gin are among the top ones in the deducted protein composition. The homology of predicted amino acids between BJ10- 2062 and other murine noroviruses is significantly higher than that with human and bovine noroviruses. Comparing tothe original strain MNV1, the 296th amino acid of BJ10-2062 VP1 capsid protein is not lysine but glutamine, which might lead to its attenuated virulence. Phylogenetic analysis grouped BJ10-2062 in genogroup V as other murine noroviruses. BJ10-2062 is in the same group with CR4 and NIH-All4 isolated from America. Conclusions Sequencing analysis of the complete genome of the murine norovirus strain BJ10-2062 from Bei- jing, China demonstrates its high homology with strains isolated from America and a set of NIH strains.
出处
《中国病毒病杂志》
CAS
2013年第5期335-342,共8页
Chinese Journal of Viral Diseases
基金
国家科技支撑计划项目(2013BAK11B01)
