《中国畜牧兽医》 ›› 2016, Vol. 43 ›› Issue (11): 2834-2843.doi: 10.16431/j.cnki.1671-7236.2016.11.005

• 生物技术 • 上一篇    下一篇

牛乳头状瘤病毒贵州株L1基因克隆及生物信息学分析

张海1, 周碧君1,2, 杨忠成3, 廖梅3, 王开功1,2, 文明1,2, 程振涛1,2, 王伟1, 冯旭芳1   

  1. 1. 贵州大学动物科学学院, 贵阳 550025;
    2. 贵州省动物疫病研究室, 贵阳 550025;
    3. 贵州省畜禽遗传资源管理站, 贵阳 550001
  • 收稿日期:2016-04-15 出版日期:2016-11-20 发布日期:2016-11-18
  • 通讯作者: 周碧君 E-mail:as.bjzhou@gzu.edu.cn
  • 作者简介:张海(1993-),男,贵州仁怀人,硕士生,研究方向:预防兽医学,E-mail:1240568761@qq.com
  • 基金资助:

    贵州省农业委员会科技专项《动物疫病防控能力提升专项》(黔农财[2015]149号);贵州省动物疫病防控与兽医公共卫生保障科技创新人才团队(黔科合人才团队[2015]4016号);贵州省黔南州社会发展科技重大计划项目《喀斯特石漠化林草牧综合治理技术研究与示范》[黔南社发科(20140325)];贵州现代肉牛产业技术体系专项(GZCYTX2015-0302-02)

Cloning and Bioinformatic Analysis of L1 Gene of BPV from Guizhou Province

ZHANG Hai1, ZHOU Bi-jun1,2, YANG Zhong-cheng3, LIAO Mei3, WANG Kai-gong1,2, WEN Ming1,2, CHENG Zhen-tao1,2, WANG Wei1, FENG Xu-fang1   

  1. 1. College of Animal Science, Guizhou University, Guiyang 550025, China;
    2. Guizhou Province Animal Disease Research, Guiyang 550025, China;
    3. Guizhou Province Station for Livestock Genetic Resources Management, Guiyang 550001, China
  • Received:2016-04-15 Online:2016-11-20 Published:2016-11-18

摘要:

为分析牛乳头状瘤病毒贵州株(BPV-GZ01株)L1基因的分子特征,预测其编码蛋白的生物学功能,本试验对BPV-GZ01株L1基因进行PCR扩增、克隆及序列测定。应用生物信息学相关软件及方法对BPV-GZ01株L1基因进行序列分析,并对其编码蛋白进行二级结构、三级结构、B细胞表位、保守结构域、跨膜结构域和信号肽预测。结果显示,L1基因序列全长为1 494 bp,编码497个氨基酸;与BPV2、BPV2-SW01、BPV2-AKS01、BPV13、BPV1株相应序列核苷酸同源性分别为99.1%、99.8%、99.4%、87.6%和82.8%,氨基酸同源性分别为98.6%、99.4%、98.4%、94.4%和91.3%;系统进化树显示,BPV-GZ01株与BPV2-SW01株亲缘关系最近;L1蛋白二级结构以无规则卷曲、β-折叠和α-螺旋区域所占比例较大,预测此蛋白可能存在6个B细胞优势抗原表位,无跨膜区域,无信号肽区域;三级结构呈弯曲状螺旋结构。本研究结果将为贵州省BPV免疫诊断及核酸疫苗研究提供理论依据。

关键词: 牛乳头状瘤病毒; L1基因; 生物信息学分析; 蛋白结构预测

Abstract:

In order to study and analyze L1 gene of bovine papillomavirus(BPV)in Guizhou province,the L1 gene of BPV-GZ01 strain was amplified,cloned and sequenced using bioinformatic softwares and methods,and the secondary structure,tertiary structure,B-cell preponderant epitope,conserved domains analysis, transmembrane domain and signal peptide of L1 gene were predicted.The results showed that the length of L1 gene was 1 494 bp,encoding 497 amino acids.The L1 gene of BPV-GZ01 strain shared an amino acid identities of 98.6%,99.4%,98.4%,94.4% and 91.3%,and a nucleotide identities of 99.1%,99.8%,99.4%,87.6% and 82.8% with those of BPV2,BPV2-SW01,BPV2-AKS01,BPV13 and BPV1 strains,respectively.The results of phylogenetic tree analysis indicated that there was a close relationship between BPV-GZ01 and BPV2-SW01 strains.The prediction of secondary structure of L1 protein indicated that the random coil,extended strand and alphahelix took a higher percentage.The L1 protein was supposed contain 6 potential antigen epitopes.And no transmembrane domains and no signal peptide were found.The tertiary structure of L1 protein was curved spiral structure.These results provided a theoretical basis for immunologic diagnosis and further research of nucleic acid vaccine of BPV.

Key words: bovine papillomavirus(BPV); L1 gene; bioinformatics analysis; protein structure prediction

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