Expression of recombinant plasmid pcDNA3.1/P12X3C with multi-genes of foot-and-mouth disease virus in BHK-21 cells

• Published in: Shengwu gongcheng xuebao Vol. 19; no. 3; p. 376
• Main Authors: Guo, Hui-Chen, Liu, Zai-Xin, Sun, Shi-Qi, Lu, Zeng-Jun, Zhou, Guang-Qing, Qi, Shu-Yun, Jin, Ye, Liu, Xiang-Tao, Xie, Qing-Ge
• Format: Journal Article
• Language: Chinese
• Published: China 01.05.2003
• Subjects: 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; Viral Proteins - metabolism
• ISSN: 1000-3061

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 ca