【作者】 赵丽荣；

【导师】 王君伟；

【作者基本信息】 东北农业大学 ， 预防兽医学， 2003， 硕士

【摘要】 鹅细小病毒(Goose Parvovirus,GPV)为小鹅瘟的病原体。小鹅瘟主要发生于1月龄内雏鹅和雏番鸭，是以急性肠炎及肝、肾、心实质脏器炎症为特征的烈性传染病，对养鹅业发展造成很大威胁。自1956年我国学者方定一首次发现并分离到GPV后，国外相继也有分离到该病毒的报道，说明该病毒在世界范围内有较为广泛的流行。 目前，对GPV的防制主要是靠疫苗接种。传统疫苗对GPV的预防和控制起到了一定的积极作用，但同时灭活苗存在生产成本高，注射局部炎症反应较强等弊端，弱毒苗又易发生毒力返强等。GPV主要免疫原性蛋白VP3的高度保守性给新型疫苗及分子诊断试剂的研制提供了思路。本研究采用脂质体转染方法，将含有完整GPV H1分离株VP3基因、报告基因LacZ、禽痘病毒早／晚期启动子LP2EP2、痘苗病毒启动子P7.5、P11和FPV-017复制非必须区的转移载体质粒pSY681VP3LacZ与FPV-017共转染鸡胚成纤维细胞，经6轮蚀斑克隆、筛选、表达，PCR鉴定和Dot-ELISA检测，证明该重组病毒已构建成功，并获得了遗传性状稳定的鹅细小病毒VP3基因的重组禽痘病毒。为新型基因工程疫苗的研制奠定了科学的基础，同时也为我国高致病力鹅细小病毒病的防治提供了物质准备。更多还原

【Abstract】 Goose Parvovirus (GPV) is the pathogen of Goslings plague. This disease is highly fatal to goslings and muscovy ducklings under 1 month of age with the characteristics of acute enteritis and hepatonephric and myocardium lesions, having made large losses to goose breeding industry. In many countries the virus has been isolated after DingYi .F firstly found and isolated it in china, which suggested that this disease has been prevalent in the world.So far as, prevention of GPV infection mainly depended on vaccines. Although the traditional vaccines have played active role in prevention and controlling the disease, many deficiencies exist still such as inactivated vaccine being expensive and inducing strongly response at the injected tissue, as well as the recovery of the attenuated vaccine. The high conservative main antigenic protein VP3 of GPV provided the access to producing the new type vaccines and diagnostic agent. In this study, the recombinant fowl-poxvirus was transfected into expressing the VP3 gene of isolated GPV H1 strain into the CEF cells with FPV-017 by liposome, which have the LacZ reporter gene, earlier / latter promoters LP2EP2 of FPV, promoters P7.5 and P7.1 of Vaccinia virus, replication unnecessary region of FPV-017. Following 6 cycles screenings, clonings, purification of blue plaques, detection of PCR and Dot-ELISA, which verified the genetic stable VP3-fowlpox virus recombinant constructed successfully. This study provided the theoretical and practical foundation for development of GPV recombinant fowl-poxvirus genetic engineering vaccine, as well as provided substance preparatory for prevention the high mortality GPV.更多还原