Construction of foot-and-mouth disease virus like particles-induced expression vectors and screening of BHK-21 cell pools.
- Author:
Shuzhen TAN
1
;
Hu DONG
1
;
Shiqi SUN
1
;
Huichen GUO
1
Author Information
- Publication Type:Journal Article
- Keywords: cell pool; foot-and-mouth disease virus (FMDV); inducible expression; plasmid construction; virus like particles (VLPs)
- MeSH: Animals; Foot-and-Mouth Disease Virus/genetics*; Capsid Proteins; Viral Proteins/metabolism*; Foot-and-Mouth Disease/prevention & control*; Tetracyclines/metabolism*; Viral Vaccines; Antibodies, Viral; Mammals/metabolism*
- From: Chinese Journal of Biotechnology 2023;39(12):4849-4860
- CountryChina
- Language:Chinese
- Abstract: Transient expression is the major method to express foot-and-mouth disease virus (FMDV) capsid proteins in mammalian cells. To achieve stable expression of FMDV capsid proteins and efficient assembly of virus like particles (VLPs) in cells, the plasmids of piggyBac (PB) transposon-constitutive expression and PB transposon-tetracycline (Tet) inducible expression vectors were constructed. The function of the plasmids was tested by fluorescent proteins. By adding antibiotics, the constitutive cell pools (C-WT, C-L127P) expressing P12A3C (WT/L127P) genes and the inducible cell pools (I-WT, I-L127P) expressing P12A3C (WT/L127P) genes were generated. The genes of green fluorescent protein, 3C protease and reverse tetracycline transactivator (rtTA) were integrated into chromosome, which was confirmed by fluorescence observation and PCR testing. The cell pool I-L127P has a stronger production capacity of capsid proteins and VLPs, which was confirmed by Western blotting and enzyme linked immunosorbent assay (ELISA), respectively. In conclusion, inducing the chromosomal expression of FMDV capsid proteins was firstly reported, which may facilitate the technical process of mammalian production of FMDV VLPs vaccine and the construction of mammalian inducible expression systems for other proteins.
