title: [Expression of recombinant plasmid pcDNA3.1/P12X3C with multi-genes of foot-and-mouth disease virus in BHK-21 cells].
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.
authors.affiliation: Chinese Academy of Agricultural Sciences, Lanzhou Veterinary Research Institute, Lanzhou 730046, China.
authors.forename: Hui-Chen
authors.initials: HC
authors.lastname: Guo
authors.forename: Zai-Xin
authors.initials: ZX
authors.lastname: Liu
authors.forename: Shi-Qi
authors.initials: SQ
authors.lastname: Sun
authors.forename: Zeng-Jun
authors.initials: ZJ
authors.lastname: Lu
authors.forename: Guang-Qing
authors.initials: GQ
authors.lastname: Zhou
authors.forename: Shu-Yun
authors.initials: SY
authors.lastname: Qi
authors.forename: Ye
authors.initials: Y
authors.lastname: Jin
authors.forename: Xiang-Tao
authors.initials: XT
authors.lastname: Liu
authors.forename: Qing-Ge
authors.initials: QG
authors.lastname: Xie
mesh_terms: 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*
chemical_list: D014764:Viral Proteins