| 猪瘟伪狂犬病重组病毒SA215(A)疫苗株的构建(初报) |
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| 引用本文： | 徐志文,郭万柱,朱玲,王晓玉.猪瘟伪狂犬病重组病毒SA215(A)疫苗株的构建(初报)[J].四川农业大学学报,2004,22(2):95-98. |
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| 作者姓名： | 徐志文 郭万柱 朱玲 王晓玉 |
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| 作者单位： | 四川农业大学,动物科技学院,四川,雅安,625014 |
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| 基金项目： | 国家“8 6 3”项目，教育部科研项目，四川省科技厅重点攻关项目，四川省教育厅重点科技攻关项目 |
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| 摘 要： | 采用磷酸钙转染系统 ,将伪狂犬病三基因缺失疫苗SA2 15株DNA与PP6 3LacZE2DNA共转染Vero细胞 ,获得SA2 15 (A) 1、SA2 15 (A) 2和SA2 15 (A) 3等 12个重组病毒株。以光生物素标记的HCVE2基因为探针进行初步鉴定后 ,挑选SA2 15 (A) 1株作BamHⅠ酶切和southern转印杂交鉴定 ,结果表明构建是成功的 ,将其命名为SA2 15(A)。直接荧光抗体检测、SDS -PAGE电泳和western免疫印迹检测结果表明HCVE2基因在重组病毒内获得表达 ,产生大小约 5 1kd的蛋白。对SA2 15 (A)株进行部分生物学特性研究 ,培养特性观察试验表明该毒株可适应Vero、BHK2 1和鸡胚成纤维细胞等多种细胞 ,但对不同细胞系表现有一定的差异。
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| 关 键 词： | 猪瘟 伪狂犬病 重组疫苗 |
| 文章编号： | 1000-2650(2004)02-0095-04 |
| 修稿时间： | 2004年6月11日 |
Construction of the Recombinant Virus Live Vaccine Strain of Pseudorabies Virus-Hog Cholera Virus |
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| XU Zhi-wen,GUO Wan-zhu,ZHU Ling,WANG Xiao-yu.Construction of the Recombinant Virus Live Vaccine Strain of Pseudorabies Virus-Hog Cholera Virus[J].Journal of Sichuan Agricultural University,2004,22(2):95-98. |
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| Authors: | XU Zhi-wen GUO Wan-zhu ZHU Ling WANG Xiao-yu |
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| Abstract: | By cotransfection of PRV Fa SA215 DNA and recombinant plasmid PP63LacZE2 DNA in Vero cells, 12 recombinant namely SA215(A)1, SA215(A)2 et al were obtained with calcium phosphate transfection system. The results obtained from biotinlabeled probe of HCV E2 indicate positive. Through digestion with BamHⅠ and Southern-blot acid hybridization of SA215(A)1 strain, this study shows the recombinant virus of PRV and HCV namely SA215(A) is successful. IFT, SDS-PAGE, electrophoresis and Western-blot confirmed HCV E2 gene is expressed in recombinant virus, about 51Kd fragment of envelope glycoprotein E2 gene. This study on the biological characteristics of SA215(A) strain shows this strain can be replicated very well on vero cells, BHK21 cells et al. There is variability of cell culture at different cell lines. SA215(A) is neutralized by positive sera of PRV, SA215(A) is of serum properties of parent virus. |
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| Keywords: | recombinant pseudorabies virus Hog Cholera virus live vector vaccine |
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