马立克氏病病毒河南分离株Meq基因的克隆与序列分析  

Cloning and phylogenetic analysis of Meq gene of Marek's disease virus isolates from Henan province

作　　者：余祖华[1,2] 滕蔓[2] 丁轲[1] 闵亚杰 禹乐乐[2] 宿靖伟[2] 程相朝[1] 罗俊[2] 

机构地区：[1]河南科技大学动物科技学院动物疫病与公共卫生重点实验室,河南洛阳471003 [2]河南省农业科学院,农业部动物免疫学重点实验室,河南省动物免疫学重点实验室,河南郑州450002 [3]洛阳普莱柯生物工程股份有限公司,河南洛阳471003

出　　处：《西北农林科技大学学报（自然科学版）》2014年第8期15-20,共6页Journal of Northwest A&F University(Natural Science Edition)

基　　金：国家自然科学基金面上项目(31072145,31372445)

摘　　要：【目的】了解马立克氏病病毒（Marek’s disease virus,MDV）河南分离株Meq基因的分子特性,为中原地区MDV流行情况提供较详细的信息,为更好地防控MD奠定基础。【方法】应用PCR方法分别扩增了17株MDV河南省野毒分离株的Meq基因片段,克隆测序后,将各毒株Meq基因序列与vv＋MDV 648A株、vvMDV RBIB株、vMDV GA株、mMDV疫苗株CVI988株和814株的Meq基因序列进行比对分析。【结果】17株分离株中除HNGS201和HNLC503的Meq基因扩增产物较预期稍大外,其余15个的片段长度为1 020bp,与预期长度相符;MDV河南分离株之间及其与参考毒株Meq基因的核苷酸同源性为99.0%～100%,部分毒株Meq基因的氨基酸序列有10个位点的氨基酸存在规律性突变;MDV河南分离株和参考株共组成3个进化分支,其中强毒株GA、RB1B和648A位于一个分支,分离株HNGS201、HNLH304、HNLC503与mMDV毒株CVI988和814位于同一分支,其余14株分离株位于另一个较大的分支。【结论】河南省可能流行着不同毒力的MDV,一些mMDV毒株的Meq蛋白存在59个或60个氨基酸的插入。【Objective】This study investigated the molecular characteristics of Meq gene of MDV isolates from Henan to provide information for the prevalence and control of MDV in Henan province.【Method】Meqgene of 17 Henan MDV isolates were amplified by polymerase chain reaction（PCR）.After cloning and sequencing,the gene sequences were compared with those of reference MDV strains including mMDVCVI988 and 814,vMDV GA,vvMDV RBIB,and vv＋MDV 648 A.【Result】PCR amplification products of15 out of the 17 isolates were consistent with expectance with the length of 1 020 bp and the isolates of HNGS201 and HNLC503had slightly longer Meqgene products.The sequences of Meqgene from different strains had the homology of 99.0%-100% with the reference MDV strains.Some isolates displayed regularly amino acid site mutations at ten positions.Phylogenetic analysis showed that the isolates and strains could be separated into three groups.The vMDV strains GA,RB1 B,and 648 A were clustered into one group,the isolates of HNGS201,HNLH304,and HNLC503 together with the mMDV strains of CVI988 and 814were clustered into the second group,and all the other 14 MDVs isolates fall into the third group.【Conclusion】The mutations site and phylogenetic analysis demonstrated that the MDV in Henan province may have different genotypes.Meq of some MDV strains had an insertion of 59 or 60amino acids.

关 键 词：马立克氏病病毒 MEQ基因 克隆 序列分析 

分 类 号：S852.65[农业科学—基础兽医学] Q78[农业科学—兽医学]