以犬2型腺病毒Ⅸ蛋白为基础的展示技术研究  

Virus Display Based on Protein Ⅸ of Canine Adenovirus Type 2

作　　者：王燕[1,2] 张守峰[2] 刘晔[2] 扈荣良[2] 张乐萃[1] 

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

出　　处：《青岛农业大学学报（自然科学版）》2008年第4期276-279,共4页Journal of Qingdao Agricultural University(Natural Science)

基　　金：国家自然科学基金(30571379)

摘　　要：以犬2型腺病毒全基因组重组质粒pPolyⅡ-CAV-2为基础,通过NcoⅠ和MluⅠ双酶切、Klenow补平,构建了不含外源启动子、大小为979bp的增强绿色荧光蛋白(EGFP)基因;通过MluⅠ和EcoRⅠ双酶切将含有犬2型腺病毒的Ⅸ蛋白的340bp的片段克隆入pEGFP-C1-dAseⅠ质粒中,构建pEGFP-SIX载体,通过AseⅠ单酶切插入EGFP,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EGFP基因,获得重组基因组质粒pPolyII-CAV-2-ⅨE(34.3kb)。酶切鉴定结果显示,目的基因均克隆进Ⅸ蛋白中。A recombinant canine adenovirus that incorporated a fusion protein of EGFP into adenovirus capsid pIX was constructed. Based on the recombinant canine adenovirus plasmid pPoly Ⅱ - CAV - 2, enhanced green fluores- cent protein （EGFP） incorporated at the minor capsid protein IX （pIX） without heterologous promoter was con- structed. The pEGFP - SIX vector that the 340bp fragment containing IX protein of canine adenovirus type 2 with Mlu I and EcoR I was inserted the pEGFP - C1 - dAse I plasmid was constructed The EGFP gene was inserted rightward between the last codon of the IX gene and the stop codon. Then we gained the recombinant plasmids that displayed EGFP gene pPoiyⅡ - CAV -2- IXE（34.3kb）. The results showed that the target genes were cloned into the IX protein by restriction enzyme digestion.

关 键 词：犬2型腺病毒 狂犬病病毒 增强绿色荧光蛋白 重组病毒 

分 类 号：S852.65[农业科学—基础兽医学]