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Chinese Journal of Biotechnology ; (12): 376-379, 2003.
Artículo en Chino | WPRIM | ID: wpr-270049

RESUMEN

In order to obtain the gene P12X3C of Foot-and-Mouth Disease Virus (FMDV) that includes full length P1, 2A, 3C and a part of 2B, the site mutation strategy was used. After being digested by Kpn I and Xba I respectively, the gene P12X3C was cloned into the pcDNA3.1 (+) expression vector. The recombinant plasmid was checked by restriction enzyme analysis and nucleic acid sequencing, and then named pcDNA3.1/P12X3C. Further, BHK-21 cells was transfected with pcDNA3.1/P12X3C by using lipoid. The proteins of Foot-and-Mouth Disease Virus, which were expressed in BHK-21 cells, were confirmed by sandwich-ELISA and fluoroscopy. The result shows the gene P12X3C is cloned into eukaryotic expression plasmid, and the recombinant eukaryotic expression plasmid pcDNA3.1/P12X3C could express proteins of Foot-and-Mouth Disease Virus in BHK-21 cells, which have immunocompetence. This study demonstrates that delivery of a recombinant eukaryotic expression plasmid containing P12X3C coding regions results in the assembly of FMDV capsid structures, which will offer experimental base to DNA vaccine of FMDV.


Asunto(s)
Animales , Cricetinae , Línea Celular , Ensayo de Inmunoadsorción Enzimática , Fluoroscopía , Virus de la Fiebre Aftosa , Genética , Vectores Genéticos , Genética , Modelos Genéticos , Plásmidos , Genética , Proteínas Virales , Genética , Metabolismo
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