›› 2015, Vol. 42 ›› Issue (1): 53-60.doi: 10.16431/j.cnki.1671-7236.2015.01.008

• 生物技术 • 上一篇    下一篇

羊口疮病毒贵州株F1L基因克隆及生物信息学分析

刘嫒1, 杨钰1, 鲜思美1,2, 刘宗胜3, 吴健4, 罗波1, 陈永翠1   

  1. 1. 贵州省动物疫病与兽医公共卫生重点实验室, 贵阳 550025;
    2. 贵州大学动物科学学院, 贵阳 550025;
    3. 毕节市动物产品质量安全监督检验所, 毕节 551700;
    4. 毕节市七星关区农牧局草地中心, 毕节 551700
  • 收稿日期:2014-08-25 出版日期:2015-01-20 发布日期:2015-02-06
  • 通讯作者: 鲜思美 E-mail:xiansimei2005@163.com
  • 作者简介:刘嫒(1991-), 女, 贵州遵义人, 硕士生, 研究方向:兽医公共卫生学, E-mail:LiuAi4735@163.com
  • 基金资助:
    贵州省科学技术基金项目[黔科合J字(2010)2260];贵州省科学技术基金项目[黔科合LH字(2014)7667];毕节市科技局项目:贵州黑山羊羔羊口疮病防治技术攻关[毕科合字(2012)24号];贵州大学"SRT计划"项目(贵大SRT字[2012]075号);贵州大学2014年大学生创新创业训练计划项目[2014(016)]

Cloning and Bioinformatic Analysis of F1L Gene of Orf Virus from Guizhou Province

LIU Ai1, YANG Yu1, XIAN Si-mei1,2, LIU Zong-sheng3, WU Jian4, LUO Bo1, CHEN Yong-cui1   

  1. 1. Key Laboratory of Animal Disease and Veterinary Public Health of Guizhou Province, Guiyang 550025, China;
    2. College of Animal Science, Guizhou University, Guiyang 550025, China;
    3. Institute of Supervision and Inspection for Animal Products Quality and Safety, Bijie 551700, China;
    4. Grassland Center of Agriculture and Animal Husbandry, Qixingguan Bureau, Bijie 551700, China
  • Received:2014-08-25 Online:2015-01-20 Published:2015-02-06

摘要: 为分析羊口疮病毒贵州株(ORFV-GZ株)F1L基因的分子特点,预测编码蛋白的生物学功能,本试验对ORFV-GZ株F1L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ORFV-GZ株F1L基因进行列分析并对其编码蛋白进行了二级结构、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,F1L基因PCR扩增产物大小为1 029 bp,编码342个氨基酸;与OV-SA00株、NZ2株、OV-IA82株和D1701株相应序列核苷酸同源性分别为98.4%、97.9%、97.8%和96.8%,氨基酸的同源性分别为98.3%、97.6%、97.3%和95.3%;系统进化树显示,ORFV-GZ株F1L基因与FJ-GT株亲缘关系最近;二级结构以α-螺旋和无规则卷曲所占比例较大,预测此蛋白可能存在7个B细胞优势抗原表位,两个跨膜区域,无信号肽区域.本试验结果将为贵州省ORFV免疫诊断及核酸疫苗研究提供理论依据.

关键词: 羊口疮病毒; F1L基因; 克隆; 生物信息学分析

Abstract: In order to study and analyze the F1L gene of Orf virus in Guizhou province (ORFV-GZ), we studied the scab materials of lambs with clinical sore mouth symptom in Guizhou province.The F1L gene was amplified, cloned and sequenced using bioinformatics softwares and methods, the secondary structure, B-cell preponderant epitope, conserved domains analysis, transmembrane domain and signal peptide of F1L gene were predicted.The results indicated the length of F1L gene was 1 029 bp, encoding 342 amino acids.The F1L gene of ORFV-GZ strain shared a nucleotide identities of 98.4%, 97.9%, 97.8% and 96.8%, and an amino acid identities of 98.3%, 97.6%, 97.3% and 95.3% with those of strains OV-SA00, NZ2, OV-IA82 and D1701, respectively.The results of phylogenetic tree analysis indicated that there was a close relationship between ORFV-GZ strain and FJ-GT.Prediction of the secondary structure of F1L indicated that the alpha helix and random coil took a higher percentage.The F1L protein was supposed contain 7 potential antigen epitopes, and two transmembrane domains, no signal peptide found.These results provided a theoretical basis for immunologic diagnosis and further research of nucleic acid vaccine of ORFV.

Key words: Orf virus (ORFV); F1L gene; clone; bioinformatics analysis

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