狂犬病毒中和线性表位重组犬腺病毒2型载体的构建  

Construction of the recombinant canine adenovirus type-2 vector containing the linear epitopes of rabies virus

作　　者：徐慧娟[1] 王晓虎[1] 李忠[1] 刘晔[2] 张守峰[2] 王颖[2] 袁子国[1] 徐振波[1] 扈荣良[2] 

机构地区：[1]吉林大学畜牧兽医学院,吉林长春130062 [2]军事医学科学院十一所流行病学研究室,吉林长春130062

出　　处：《畜牧与兽医》2008年第6期1-4,共4页Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金(30471294)

摘　　要：本试验成功构建了表达狂犬病病毒糖蛋白膜外区3个中和线性表位的重组犬腺病毒2型两个重组质粒。引用相关文献已报道的狂犬病病毒糖蛋白膜外区的3个中和线性表位基因,通过有柔性氨基酸将其3个表位基因相连。将重叠延伸PCR获得3个串联表位基因片段克隆入含犬腺病毒2型YCA18株纤突头部HI环的中间载体pBluescript II KS(+)中,然后用SalⅠ和PacⅠ将中间载体中含3个串联表位的纤突部分回收,克隆至缺失纤突部分的犬腺病毒2型全基因组重组质粒pPloyII-CAV-2及pPloyII-CAV-2-CGS中。经鉴定成功获得在犬腺病毒2型纤突头部HI环处插有狂犬病毒3个串联表位的两个重组质粒,分别命名为pPoly II-CAV-2-RG和pPoly II-CAV-2-CGS-RG。经软件对犬腺病毒2型纤突蛋白的构象分析表明,在其HI环插入外源抗原表位基因不影响其免疫原性。本试验为进一步研制多抗原表位展示性重组疫苗、新型狂犬病疫苗奠定基础。DNA fragment of three linear epitopes of rabies virus glycoprotein was amplified by overlap extension PCR and cloned into middle vector pBluescript ⅡKS （ ＋ ） containing the HI loop of canine adenovirus type-2 strain YCA18. The fragment with spike and linear epitopes was obtained by Sal Ⅰ and Pac Ⅰ digestion from the middle vector, and then cloned into the recombinant plasimds-pPloyⅡ-CAV-2 and pPloyⅡ-CAV-2-CGS with the total genome of canine adenovirus type-2 which was lacked of spike epitope. Two recombinant canine adenovirus type-2 vectors containing three linear epitopes of rabies virus glycoprotein, pPolyⅡ-CAV-2-RG and pPoly Ⅱ-CAV-2-CGS-RG, were successfully constructed. The conformational analysis revealed that immunogenicity of spike protein was not affected after the gene fragment of three linear epitopes from rabies virus glycoprotein was inserted into the spike gene of canine adenovirus type-2. The two recombinant canine adenovirus type-2 vectors are background for the studies on recombinant vaccines of displaying several epitopes and a new type vaccine against rabies.

关 键 词：狂犬病毒 糖蛋白 线性表位 犬腺病毒2型 

分 类 号：S852.65[农业科学—基础兽医学]