展示狂犬病病毒保护性抗原的重组犬2型腺病毒的构建  

Construction of a recombinant canine adenovirus displaying the protective antigen of rabies virus

作　　者：王燕[1] 张守峰[1] 崔艳[1] 徐振波[1] 于恒智[2] 扈荣良[1] 张乐萃[2] 

机构地区：[1]军事医学科学院军事兽医研究所,长春130062 [2]青岛农业大学动物科技学院,青岛266109

出　　处：《中国动物检疫》2008年第2期24-26,共3页China Animal Health Inspection

基　　金：国家自然科学基金(30571379)

摘　　要：本试验以犬2型腺病毒全基因组重组质粒pPolyⅡ-CAV-2为基础,以8202株狂犬病病毒基因组为模板,通过PCR扩增得到狂犬病病毒糖蛋白膜外区基因(8202-G″)。通过MluⅠ和EcoRⅠ双酶切pPolyⅡ-CAV-2质粒,获得含有Ⅸ蛋白的340bp片段,将其克隆入pEGFP-C1-dAseⅠ质粒中,构建pEGFP-SIX载体,该载体经AseⅠ单酶切插入8202-G″,即在编码Ⅸ蛋白基因的最后密码子与终止密码子之间按与Ⅸ蛋白编码链相同转录方向插入8202-G″基因,获得重组基因组质粒和pPolyII-CAV-2-ⅨG(34.8kb)。酶切鉴定结果显示,目的基因均克隆进Ⅸ蛋白中。An recombinant genome of canine adenovirus type-2 was constructed in this experiment. The external part of glycoprotein was amplified through RT-PCR and cloned into Ase I site of pEGFP-C 1-dAse I containing the capsid protein IX of canine adenovirus type-2, pEGFP-SIX was obtained. The capsid protein IX of CAV-2 genome was replaced with the recombinant protein IX that was released by Ase I from pEGFP-SIX rightward between the last codon of the IX gene and the stop codon. As a result, the recombinant genome of CAV-2 was obtained. The results showed that the target genes were cloned into the IX protein by restriction enzyme digestion.

关 键 词：狂犬病 糖蛋白 犬2型腺病毒 重组病毒 

分 类 号：S852.655[农业科学—基础兽医学]