| NDV长春株和四平株HN/F核酸疫苗的构建及表达 |
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| 引用本文： | 龚伟,金宁一,薛立娟,罗琴芳,孙大辉,葛涛,李萍.NDV长春株和四平株HN/F核酸疫苗的构建及表达[J].中国兽医学报,2002,22(2):105-107. |
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| 作者姓名： | 龚伟 金宁一 薛立娟 罗琴芳 孙大辉 葛涛 李萍 |
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| 作者单位： | 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 |
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| 基金项目： | 国家“973”资助项目 ( G1990 1190 2 ) |
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| 摘 要： | 将新城疫病毒（NDV）长春株和四平株HN插入pIRES1多克隆位点（EcoRⅤ）中，构建成核酸表达疫苗pIRcHN和pIRsHN，然后切除pIRcHN和pIRsHN的新霉素基因，将长春株和四平株F基因分辊手稿其中，构建成pIRcHNF和pIRsHNF，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pIRcHN最高，pIRsHN次之，pIRcHNF和pIRsHNF较低。将重组疫苗转染Hela细胞，用兔抗鸡IgY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。
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| 关 键 词： | NDV HN基因 F基因 核酸疫苗 新城疫病毒 |
| 文章编号： | 1005-4545(2002)02-0105-03 |
| 修稿时间： | 2000年12月20 |
Construction and Expression of NDV HN/F genes DNA Vaccine Expresssion Plasmids |
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| GONG Wei,JIN Ning-yi,XUE Li-juan,LUO Qin-fang,SUN Da-hui,GE Tao,LI Ping.Construction and Expression of NDV HN/F genes DNA Vaccine Expresssion Plasmids[J].Chinese Journal of Veterinary Science,2002,22(2):105-107. |
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| Authors: | GONG Wei JIN Ning-yi XUE Li-juan LUO Qin-fang SUN Da-hui GE Tao LI Ping |
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| Abstract: | 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. |
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| Keywords: | NDV HN gene F gene nucleic acid vaccine |
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