猪细小病毒自然弱毒N株VP1基因的扩增与序列分析  

Amplification and Sequence Analysis of VP1 Gene of Porcine Parvovirus N Strain

作　　者：李斌[1] 赵武[1] 梁家幸[1] 梁保忠[1] 白安斌[1] 姚瑞英[1] 黄安国[1] 何颖[1] 蒋玉雯[1] 

机构地区：[1]广西兽医研究所,广西南宁530001

出　　处：《动物医学进展》2008年第10期5-9,共5页Progress In Veterinary Medicine

基　　金：广西科学基金项目(桂科自0728102)

摘　　要：对猪细小病毒自然弱毒N株(PPV-N株)VP1基因扩增及测序,利用生物信息学技术预测VP1蛋白的二级结构与B细胞抗原表位,以及分析该蛋白的氨基酸差异位点及密码子偏爱性。结果表明,PPV-N株VP1基因与弱毒参考毒株NADL-2VP1基因同源性最高,亲缘关系最近。PPV-N株VP1蛋白的二级结构较为复杂,以β-折叠为主,并有较多的转角和无规则卷曲区域,在序列的20～56、61～80、86～130、136～188区域为B细胞抗原表位的优势区域。PPV强、弱毒株VP1蛋白存在5个氨基酸差异位点(T-365-I,G-528-D,Q-533-H,P-586-S,K-715-R),这些位点处氨基酸残基抗原性与毒力成正相关。PPV-N株VP1蛋白在A、C、E、G、H、I、K、N、Q、R、S、T、V和Y氨基酸于密码子选择上存在一定的偏爱性。The VP1 gene of porcine parvovirus N strain （PPV-N strain） was amplified and sequenced. Then the secondary structure of VP1 protein was predicted by the methods of Chou-Fasman, Gamier-Robson and Karplus-Schultz based on PPV-N strain VP1 gene. Hydrophilicity, surface probability and antigenic index were acquired by the methods of Kyte-Doolittle, Emini and Jameson-Wolf, respectively. Combining these results, the B cell epitopes for VP1 protein of PPV-N strain were predicted. The varying sites of amino acid and eodon preference of VP1 protein of PPV-N strain were also analysed by the methods of Clustal W and EMBOSS, respectively. The results showed that the mutation of PPV-N strain VP1 gene was tiny, PPV-N strain and NADL-2 strain from Canada were classified as the natural attenuated porcine parvovirus strain, β-sheet regions were the main conformation in VP1 protein which had much turn regions and coil regions, and the predominant B cell epitopes were probably in the regions of 20-56,61-80,86-130 and 136-188. The varying sites of amino acid between PPV Virulent strain and PPV attenuated strain were T-365-I, G-528-D, Q-533-H, P-586-S, K-715-R, and the antigenieity of these amino acid sites showed positive correlation with the virulence of PPV. Codon preference of PPV-N strain VP1 protein were infered in A,G,E,G,H,I,K,N,Q,R,S,T,V and Y amino acid. This study would be useful for research of natural attenuated virulence mechanism and reverse vaccine of PPV-N strain.

关 键 词：PPV—N株 VP1基因 扩增 生物信息学分析 

分 类 号：S852.659.2[农业科学—基础兽医学] S858.28[农业科学—兽医学]