[表达口蹄疫病毒VP1基因的重组牛疱疹病毒1型的构建与鉴定]
[Construction and identification of recombinant BHV-1 expressing foot and mouth disease virus VP1 gene].
作者信息
Ren Xiangang, Xue Fei, Zhu Yuanmao, Tong Guanzhi, Wang Yanhui, Feng Junke, Zu Lichuang, Li Jiao, Shi Hongfei, Gao Yuran
机构信息
Division of Livestock Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin 150001, China.
出版信息
Wei Sheng Wu Xue Bao. 2009 May;49(5):677-82.
OBJECTIVE
In order to construct the recombinant bovine hepervirus-1 (BHV-1) which expressed foot and mouth disease virus (FMDV) VP1 gene, we constructed a BHV-1 gE gene transfer vector by inserting the synthetic VP1 gene of FMDV (O/China/99) under the immediate-early promoter of cytomegalovirus.
METHODS
The mixtures of parental virus (BHV-1/gE(-)/LacZ+) DNA and transfer vector was transfected into bovine turbinate cells using calcium phosphate-mediated transfection. Then the propagated viruses were harvested. The recombinant BHV-1 (designated BHV-1/gE(-)/VP1) was obtained by selection for white virus plaques.
RESULTS
PCR results showed that VP1 gene was successfully inserted into the genome of BHV-1/gE(-). The expression of VP1 in infected cells was proved by indirect immunofluorescence assay and Western blotting.
CONCLUSION
The research provided a basis for development of BHV-1 vector vaccines for FMD and other important bovine infectious diseases.
目的
为构建表达口蹄疫病毒（FMDV）VP1基因的重组牛疱疹病毒1型（BHV-1），我们通过将FMDV（O/China/99）的合成VP1基因插入巨细胞病毒的立即早期启动子下，构建了一个BHV-1 gE基因转移载体。
方法
使用磷酸钙介导的转染方法，将亲本病毒（BHV-1/gE(-)/LacZ+）DNA与转移载体的混合物转染到牛鼻甲细胞中。然后收获增殖的病毒。通过选择白色病毒斑获得重组BHV-1（命名为BHV-1/gE(-)/VP1）。
结果
PCR结果表明VP1基因成功插入BHV-1/gE(-)的基因组中。间接免疫荧光试验和蛋白质印迹法证实了VP1在感染细胞中的表达。
结论
该研究为开发用于口蹄疫和其他重要牛传染病的BHV-1载体疫苗提供了依据。
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