Expression of recombinant plasmid pcDNA3.1/P12X3C with multi-genes of foot-and-mouth disease virus in BHK-21 cells / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 376-379, 2003.
Article
in Chinese
| WPRIM
| ID: wpr-270049
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.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Plasmids
/
Viral Proteins
/
Enzyme-Linked Immunosorbent Assay
/
Fluoroscopy
/
Cell Line
/
Foot-and-Mouth Disease Virus
/
Genetic Vectors
/
Genetics
/
Metabolism
/
Models, Genetic
Limits:
Animals
Language:
Chinese
Journal:
Chinese Journal of Biotechnology
Year:
2003
Type:
Article
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