中文摘要：

为评价重组腺病毒融合表达猪瘟病毒（CSFV）E0和E2蛋白的免疫保护效果，本研究以CSFV基因组为馍板．应用RT—PCR扩增E0和E2蛋白的编码基因，通过pET-32a载体将E0和E2基因串联，形成pET—E0-E2重组质粒用KpnI和NotI双酶切pET—E0-E2得到E0-E2融合基因，定向亚克隆于宇梭载体pAdTrack—CMV中，采用“两步转化法”在细菌内同源重组，构建携带E0-E2基因的重组腺病毒转移载体质粒pAdEasy—E0-E2，经PacI酶切线性化后转染人胚胎肾细,Ig（HEK293），成功包装出重组腺病毒（rAd—E0-E2），PCR和westernblot捡测表明，E0-E2基因已重组于昧病毒基因组中并获得表达将rAd—E0-E2接种于小鼠和猪，并通过ELISA进行抗体检测另外，将tAd—E0．E2经肌肉2次（间隔7d）免疫接种6周龄～7周龄猪，3周后用10 3TCID50 CSFV石门株攻毒结果表明，rAd-E0-E2免疫组6／7头存活，各组织器官带毒时间不超过10d，而非重组腺病毒rAd—CMV免疫组和空白对照组全部死亡，各组织器官均能分离到CSFV，结果提示，rAd—E0-E2能使免疫猪抵抗CSFV强毒攻击，为进一步研制猪瘟基因工程疫苗提供了实验依据．

英文摘要：

A recombinant adenovirus rAd-E0-E2 fusion-expressing the E0 and E2 gene of atypical classical swine fever virus （CSFV） was constructed by cloning CSFV E0 and E2 genes into the adenoviral shuttle plasmid pAdTrack-CMV and transfected into 293 cells. The integration of E0-E2 gene in the recombinant adenovirus genome was confirmed by PCR, and the expression of E0-E2 protein in 293 cells was detected by western blot. Pigs were administered with rAd-E0-E2 intramascularly two times at a 7 days interval followed by challenge with 10 3 TCID50 of CSFV Shimen strain 3 weeks later. The results showed that 6/7 of the pigs vaccinated with rAd-E0-E2 were protected, and the persistence of CSFV in organs was less than 10 days. None of the pigs vaccinated with the human adenovirus （rAd-CMV） and DMEM survived. These results demonstrated that the recombinant adenovirus rAd-E0-E2 constructed in this study could induce effective protection in pigs against CSFV challenge, which provided a reference for the development of CSF genetic engineering vaccine.