伪狂犬病病毒Min-A株gD基因的克隆及其真核表达质粒的构建  

Cloning and sequencing of the gD gene of pseudorabies virus Min-A strain

作　　者：胡涛[1] 崔保安[1] 王岩 祁小乐[1] 王丽娟[1] 

机构地区：[1]河南农业大学牧医工程学院,河南郑州450002 [2]郑州牧业高等专科学校,河南郑州450008

出　　处：《中国预防兽医学报》2004年第1期25-27,共3页Chinese Journal of Preventive Veterinary Medicine

摘　　要：参考GeneBank收录的伪狂犬病病毒gD基因的序列设计了一对引物,对PRVMin＿A株进行了PCR扩增,扩增产物克隆于pGEM＿TEasy载体。对重组质粒PGTE＿gD进行限制性内切酶分析和基因测序,证实了克隆片段的可靠性。测序结果表明目的片段包含一个1203bp的开放性阅读框(ORF),编码400个氨基酸组成的多肽。将gD基因亚克隆至真核表达载体pcDVA3.1＿的CMV启动子下游,构建了真核表达质粒pcDNA＿gD,为下一步基因免疫奠定了基础。A pair of primers were designed according to the sequences pulished by the GenBank in order to amplifiy gD gene of the pseudorabies virus Min-A strain.The gD gene were obtained by polymerase chain reaction(PCR), and then cloned into the pGEM-T Easy vector. The recombinant plasmid of pGTE-gD was identified by restriction enzyme analysis and sequencing ,Which proved completely its validity. Nucleotide sequencing revealed that this fragment contained an open reading frame of 1203 bp encoding a 400 aa protein. The gD geng is subcloned into pcDNA3.1-,an eukaryotic expressing vector.The constructed expressing plasmid pcDNA-gD will be used for future gene vaccination.

关 键 词：伪狂犬病病毒 Min-A菌株 GD基因 基因克隆 真核表达质粒 双链DNA 

分 类 号：S852.659.1[农业科学—基础兽医学] Q785[农业科学—兽医学]