NDV长春株和四平株HN/F核酸疫苗的构建及表达  

Construction and Expression of NDV HN/F genes DNA Vaccine Expresssion Plasmids

作　　者：龚伟[1] 金宁一[1] 薛立娟[1] 罗琴芳[1] 孙大辉[1] 葛涛[1] 李萍[1] 

机构地区：[1]解放军军需大学军事兽医研究所解放军基因工程重点实验室,吉林长春130062

出　　处：《中国兽医学报》2002年第2期105-107,共3页Chinese Journal of Veterinary Science

基　　金：国家"973"资助项目 ( G1990 1190 2 )

摘　　要：将新城疫病毒 (NDV)长春株和四平株 HN插入 p IRES1多克隆位点 (Eco R )中 ,构建成核酸表达疫苗 p IRc HN和 p IRs HN,然后切除 p IRc HN和 p IRs HN的新霉素基因 ,将长春株和四平株 F基因分别插入其中 ,构建成 p IRc HNF和 p IRs HNF,而后分别转染 Hela细胞。经血凝效价测定、Western blot分析 ,弱毒株构建的核酸疫苗的血凝活性比强毒株高 1个数量级 ,Hela细胞表达的 HN蛋白量以 p IRc HN最高 ,p IRs HN次之 ,p IRc HNF和 p IRs HNF较低。将重组疫苗转染 Hela细胞 ,用兔抗鸡 Ig Y进行间接免疫荧光试验 ,结果 ,在细胞膜和细胞浆中观察到了特异性的黄绿色荧光 ,证明表达产物具有特异性。Two nucleic acid vaccine plasmids, pIRcHN and pIRsHN were constructed by inserting HN of NDV Changchun or Siping strain into multicloning site of pIRES1neo. Another two nucleic acids, pIRcHNF and pIRsHNF that coexpressing the HN and the F were constructed by inserting the F gene of NDV Changchun or Siping strain into pIRcHN and pIRsHN. The expression of HN in Hela cells was confirmed by hemagglutination test, Western blotting and indirect immunoflurescence. The results showed that the hemagglutination titer of pIRcHN and pIRsHN were two fold higher than that of pIRcHNF and pIRsHNF. The expression product of pIRcHN had more purpose protein than the other three plasmids. Specific bright yellow green flurescene was observed around the membrane and across the plasmas of the transfected cells when tested with anti-chicken IgY, FITC conjugate.

关 键 词：NDV HN基因 F基因 核酸疫苗 新城疫病毒 

分 类 号：S852.65[农业科学—基础兽医学] Q78[农业科学—兽医学]