中文摘要：

根据马动脉炎病毒（Equine arteritis virus，EAV）Bucyrus株全基因组序列设计并合成EAV特异引物，进而应用RT—PCR技术分6段扩增了EAV全基因组cDNA。将扩增的各个cDNA重叠片段PE124、PE631、PE1854、PE5191、PE61107、PE97Q分别克隆到载体PCR BluntⅡ-TOPO中，建立了EAV初级cDNA克隆。在扩增5′末端时，引入NotⅠ酶切位点和T3启动子序列；在基因组3′末段PolyA尾引入XhoⅠ酶切位点，后者供cDNA模板的线性化之用。将扩增片段在PCR BluntⅡ-TOPO中依次连接，最后亚克隆到质粒pBluescriptⅡKS（＋）中，获得了EAV全长基因组cDNA克隆pWEAV。核酸序列分析表明：该毒株基因组全长为12704个核苷酸，与EAV Bucyrus分离株的同源性为99．1％；全长基因组中有104个核苷酸发生突变，相应地导致编码区内34个氨基酸的改变。

英文摘要：

To develop a reverse genetics system of equine arteritis virus （EAV）, six pairs of oligonueleotides were designed based on the full-length genomie sequence of EAV Bucyrus strain. Using RT-PCR technique,six overlapping eDNA fragments, designated as PE124, PE631, PE1854, PE5191, PE61107 and PE97Q, respectively, were amplified, followed by cloning into pCR Blunt Ⅱ-TOPO vector. A Not Ⅰ site and a T3 promoter were introduced immediately upstream of 5′-end,while a Xho Ⅰ Site was engineered downstream of 3′end of EAV poly（A） tail. Using pBluescript Ⅱ KS（＋） as a plasmid vector,the full length eDNA clone pWEAV of Bucyrus strain was obtained by connecting the six eDNA fragments utilizing single restriction endonuclase site. The construction of a full length genomic cDNA clone of EAV is a crucial step to obtain the infectious clone,which may facilitate further dissecting of structure and function relationship of EAV genome.