鸭瘟病毒gC基因的克隆及其分子特性分析  

Cloning and molecular characterization of gC gene of duck plague virus

作　　者：徐超[1] 李欣然[1] 信洪一[1] 练蓓[1] 程安春[1,2] 汪铭书[1,2] 朱德康[1,2] 贾仁勇[2] 罗启慧[2] 陈孝跃[1] 

机构地区：[1]四川农业大学动物医学院禽病防治研究中心,四川雅安625014 [2]动物疫病与人类健康四川省重点实验室,四川雅安625014

出　　处：《中国兽医科学》2008年第12期1038-1044,共7页Chinese Veterinary Science

基　　金：国家"十一五"科技支撑计划项目(2007Z06-017);国家自然科学基金项目(30771598);教育部"新世纪优秀人才支持计划"项目(NCET-06-0818);高等学校科技创新工程重大项目培育资金项目(706050);四川省基础研究重大项目(05JY029-109/05JY029-003/07JY029-016/07JY029-017);四川省重点建设学科项目(SZD0418)

摘　　要：对已构建的鸭瘟病毒(DPV)DNA文库重组质粒测序后,运用ORF Finder、BLASTN等工具进行全面生物信息学分析,根据分析结果设计引物,以DPV DNA为模板进行PCR扩增和T克隆,获得完整基因,经测序和探针核酸斑点杂交进行验证。结果证实,获得的完整ORF片段(ORF1296)为DPV的gC基因(GenBank登录号EU076811),全长1 296 bp,与已报道的其他疱疹病毒gC基因具有较高的同源性,它编码432个氨基酸,蛋白分子质量为45 ku、等电点(PI)为5.292,具有疱疹病毒典型I型膜蛋白结构特征。gC基因进化树分析表明,DPV与α-疱疹病毒亚科的鸟源疱疹病毒亲缘关系最近,编码的gC蛋白多为柔性区域,富含无规卷曲,含少量α-螺旋和β-折叠,氨基酸序列的第21～24、50～58、67～70、161～171、273～281、387～393位最有可能为抗原表位。此结果为进一步开展DPV gC基因及其编码蛋白的深入研究提供了有用数据。A new gene（gC） from duck plaque virus（DPV） DNA gene library constructed in Key Laboratory of Animal Disease and Human Health of Sichuan Province was selected and sequenced. A complete open reading frame（ORF1296）was found by bioinformatics tools of ORF Finder and BLASTN. The gene was identified again by PCR, sequencing and nucleic acid dot-hybridization and belonged to DPV DNA, which shared high homology with the other herpesvirus gC gene （GenBank Accession No. EU076811）. Bioinformatics analysis showed that the ORF of 1 296 bp encoding gC gene and encoding 432 amino acids was obtained. The protein molecular weight was 45 ku,and its isoelectric point（PI） was 5. 292,which possessed the typical characterization of type Ⅰ membrane protein from family herpesviridae. The phylogenetic analysis showed that the gC gene had a closest evolutionary relationship with the gallid herpesvirus and the protein had many flexible regions and random coils, but only a few of α-helices and β-strands. The positions 21 to 24,50 to 58,67 to 70,161 to 171,273 to 281 and 387 to 393 of the amino acid sequences were most potential antigenic epitopes. The results might provide foundation for further studies of DPV gC.

关 键 词：鸭瘟病毒 GC基因 克隆 分子特性 

分 类 号：S852.659.1[农业科学—基础兽医学] Q786[农业科学—兽医学]