狂犬病病毒核蛋白基因重组犬2型腺病毒的构建研究  

Construction and identification of a rabies virus nucleoprotein-canine adenovirus recombinant

作　　者：李海涛[1] 扈荣良[1] 张守峰[1] 肖跃强[1] 袁慧君[1] 涂长春[1] 

机构地区：[1]军事医学科学院军事兽医研究所病毒学研究室,长春130062

出　　处：《免疫学杂志》2005年第6期453-456,共4页Immunological Journal

基　　金：国家自然科学基金项目(30170704);国家"863"计划项目(2004AA213101)资助

摘　　要：目的获得具有感染性的狂犬病病毒核蛋白犬2型腺病毒,并对重组病毒生物学和免疫学特性进行初步探讨。方法对克隆的犬2型腺病毒全基因组E3区进行部分缺失后,插入由CMV启动子、狂犬病病毒SRV9株核蛋白基因和牛生长激素polyA构成的长2392bp的表达盒,然后通过AscⅠ和PvuⅠ双酶切,游离重组全基因组,转染犬肾细胞系。结果盲传5代后,细胞产生典型病变。经鉴定,确定得到了狂犬病病毒核蛋白重组犬2型腺病毒(CAV2RN)。Westernblot检测表明,狂犬病病毒核蛋白在重组犬2型腺病毒中得到了表达。结论重组病毒可以诱导受试犬产生针对犬2型腺病毒和狂犬病病毒核蛋白的特异性抗体。Objective To construct a rabies virus nucleoprotein containing canine adenovirus type 2 （CAV-2） recombinant, and to evaluate the biological and immunological properties of the recombinant virus. Methods The expression cassette of nucleoprotein consisted of the human cytomegalovirus （CMV） immediate early promoter, rabies virus strain SRV9 nucleoprotein cDNA, and polyadenylation signal of the bovine growth hormone. By using two Ssp Ⅰ sites in the E3 region of CAV-2 strain YCA18, a DNA fragment of 859 bp was replaced by the 2 392 bp expression cassette of the nucleoprotein. The recombinant genome was released by Asc Ⅰ and Pvu Ⅰ double digestion, then and was transfected into MDCK cells by LipofectamineTM reagent. Results Typical cytopathogenic effect was observed 10 d post-transfection. SDSPAGE and Western blotting analysis showed that the nucleoprotein was expressed. Conclusion The recombinant virus could stimulate specific immune responses to CAV-2 and rabies virus.

关 键 词：狂犬病病毒 核蛋白 犬2型腺病毒 重组病毒 

分 类 号：R373[医药卫生—病原生物学]