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| 鸭瘟病毒CHv株UL34基因的克隆及分子特性分析 | |
| 引用本文： | 钟小容,程安春,汪铭书,朱德康,龚永强,贾仁勇,罗启慧,刘菲,陈孝跃.鸭瘟病毒CHv株UL34基因的克隆及分子特性分析[J].中国兽医学报,2010,30(2). |
| 作者姓名： | 钟小容 程安春 汪铭书 朱德康 龚永强 贾仁勇 罗启慧 刘菲 陈孝跃 |
| 作者单位： | 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;动物疫病与人类健康四川省重点实验室重庆市合川区环境监测站,重庆,401520 2. 四川农业大学,动物医学院,禽病防治研究中心,四川,雅安,625014;动物疫病与人类健康四川省重点实验室 3. 四川农业大学,动物医学院,禽病防治研究中心,四川,雅安,625014;重庆市合川区动物监督所,重庆,401520 4. 动物疫病与人类健康四川省重点实验室 5. 四川农业大学,动物医学院,禽病防治研究中心,四川,雅安,625014动物疫病与人类健康四川省重点实验室 |
| 基金项目： | 教育部长江学者和创新团队发展计划创新团队资助项目，现代农业产业技术体系建设专项资金资助项目 |
| 摘 要： | 通过生物信息学分析、斑点杂交实验、克隆和测序证实了本实验室构建鸭瘟病毒(Duck plague virus,DPV)CHv株基因文库时新分离的基因(GenBank登录号EF643562)为DPV UL34基因,进一步运用有关分子生物学软件对该基因进行了分子特性分析。结果表明,该基因大小为831bp,编码276个氨基酸的多肽,含有疱疹病毒UL34类似蛋白家族保守区。系统进化树分析显示,其与α疱疹病毒亚科中马立克病毒属的GaHV-2,GaHV-3,MeHV-1,EHV-1和水痘病毒属的CeHV-9,HHV-3的亲缘关系最近,揭示DPV-CHv株应该被划为α疱疹病毒亚科中的单独一类。该编码蛋白在252～274位氨基酸有一个明显的跨膜区,C末端有一微体靶向序列(ARL),但无信号肽;含有4个酪蛋白激酶Ⅱ磷酸化位点,3个蛋白激酶C磷酸化位点,5个N-豆蔻酰化位点和1个N-糖基化位点;亚细胞定位主要位于细胞核;密码子偏爱性分析显示其偏爱的密码子第3位碱基多以A和T结尾,属于低表达基因;不同密码子使用频率与酵母、大肠杆菌和人的密码子使用频率比较表明,其与大肠杆菌和酵母较接近,与人差异较大。该分析结果揭示DPV UL34基因为一C端锚定的Ⅱ型膜蛋白基因,体外表达选择大肠杆菌和酵母表达系统较为合适,同时该结果也为进一步开展DPV UL34基因的生物学功能研究具有一定的参考作用。 |
| 关 键 词： | 鸭瘟病毒 UL34基因 分子特性 |
Cloning and molecular characteristic analysis of UL34 gene from duck plague vi-rus CHv strain |
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| ZHONG Xiao-rong,CHENG An-chun,WANG Ming-shu,ZHU De-kang,GONG Yong-qiang,JIA Ren-yong,LUO Qi-hui,LIU Fei,CHEN Xiao-yue.Cloning and molecular characteristic analysis of UL34 gene from duck plague vi-rus CHv strain[J].Chinese Journal of Veterinary Science,2010,30(2). | |
| Authors: | ZHONG Xiao-rong CHENG An-chun WANG Ming-shu ZHU De-kang GONG Yong-qiang JIA Ren-yong LUO Qi-hui LIU Fei CHEN Xiao-yue |
| Abstract: | A 831-bp complete open reading frame (ORF) of the duck plague virus UL34 gene (GenBank accession No: EF643562) was isolated in our laboratory by constructing the genomic library of DPV CHv strain. This gene was identified by NCBI ORF finder, dot-blot hybridization, cloning and sequencing. A large number of bioinformatics soft-ware was then used to analyze its molecular characteristics. The results indicated that this gene encodes an estimated 276 putative protein,contains the conserved domain of the Herpesvirus UL34-1ike protein. Phylogenetic tree based on the amino acids sequences showed DPV has a close evolutionary relationship with GaHV-2, GaHV-3, MeHV-1, EHV1,CeHV-9 and H HV3,which classified into the Mardivirus or Varicellovirus ,indicating that the DPV should be placed into a single cluster within the subfamily Alphaherpes.virinae. An obvious transmembrane region was loca-ted between 252-274 amino acids and also has a microbody targeting signal in the C terminal,but without a signal peptide. Four casein kinase Ⅱ phosphorylation sites, four protein kinase C phosphorylation sites, five N-myristoyla-tion sites and one N-glycosylation site were searched in the protein. Most of the total proteins probably situated in the nucleus. The codon usage patterns of the UL34 gene in DPV showed it was a low expression gene with AT-riched at the third position. These results strongly suggested that the UL34 protein of DPV-CHv strain was a tail-anchored type Ⅱ membrane protein,and the expression in Escherichia coli or yeast maybe the better,as well as pro-vide rational and valuable data to elucidate biological function of the gene. |
| Keywords: | duck plague virus UL34 gene molecular characteristic |
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