[Expression of recombinant plasmid pcDNA3.1/P12X3C with multi-genes of foot-and-mouth disease virus in BHK-21 cells].
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.
Guo
Hui-Chen
HC
Chinese Academy of Agricultural Sciences, Lanzhou Veterinary Research Institute, Lanzhou 730046, China.
Liu
Zai-Xin
ZX
Sun
Shi-Qi
SQ
Lu
Zeng-Jun
ZJ
Zhou
Guang-Qing
GQ
Qi
Shu-Yun
SY
Jin
Ye
Y
Liu
Xiang-Tao
XT
Xie
Qing-Ge
QG
D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D004797:Enzyme-Linked Immunosorbent Assay; D005471:Fluoroscopy; D029544:Foot-and-Mouth Disease Virus / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics; D008957:Models, Genetic; D010957:Plasmids / Q000235:genetics; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*
D014764:Viral Proteins