Expression of recombinant plasmid pcDNA3.1/P12X3C with multi-genes of foot-and-mouth disease virus in BHK-21 cells.
- Author:
Hui-Chen GUO
1
;
Zai-Xin LIU
;
Shi-Qi SUN
;
Zeng-Jun LU
;
Guang-Qing ZHOU
;
Shu-Yun QI
;
Ye JIN
;
Xiang-Tao LIU
;
Qing-Ge XIE
Author Information
1. Chinese Academy of Agricultural Sciences, Lanzhou Veterinary Research Institute, Lanzhou 730046, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Cell Line;
Cricetinae;
Enzyme-Linked Immunosorbent Assay;
Fluoroscopy;
Foot-and-Mouth Disease Virus;
genetics;
Genetic Vectors;
genetics;
Models, Genetic;
Plasmids;
genetics;
Viral Proteins;
genetics;
metabolism
- From:
Chinese Journal of Biotechnology
2003;19(3):376-379
- CountryChina
- Language:Chinese
-
Abstract:
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.
