Co-expression, purification and bioassay of three avian viral antigens.
- Author:
Suling ZHANG
1
;
Mengyue WANG
1
;
Yanwei WANG
1
;
Peng WU
1
;
Wenqiang PANG
1
;
Kegong TIAN
1
Author Information
- Publication Type:Journal Article
- Keywords: EDSV Fiber protein; FAdV-4 Fiber-2 protein; IBDV VP2 protein; co-expression; codon optimization; promoter optimization
- MeSH: Animals; Antigens, Viral/genetics*; Biological Assay; Chickens/immunology*; Escherichia coli/genetics*; Infectious bursal disease virus/immunology*; Poultry Diseases; Vaccines, Synthetic/isolation & purification*; Viral Structural Proteins/immunology*; Viral Vaccines/immunology*
- From: Chinese Journal of Biotechnology 2020;36(10):2066-2075
- CountryChina
- Language:Chinese
- Abstract: To achieve uniform soluble expression of multiple proteins in the same Escherichia coli strain, and simplify the process steps of antigen production in genetic engineering subunit multivalent vaccine, we co-expressed three avian virus proteins including the fowl adenovirus serotype 4 (FAdV-4) Fiber-2 protein, infectious bursal disease virus (IBDV) VP2 protein and egg-drop syndrome virus (EDSV) Fiber protein in E. coli BL21(DE3) cells after optimization of gene codon, promoter, and tandem expression order. The purified proteins were analyzed by Western blotting and agar gel precipitation (AGP). The content of the three proteins were well-proportioned after co-expression and the purity of the purified proteins were more than 80%. Western blotting analysis and AGP experiment results show that all the three co-expression proteins had immunoreactivity and antigenicity. It is the first time to achieve the three different avian virus antigens co-expression and co-purification, which simplified the process of antigen production and laid a foundation for the development of genetic engineering subunit multivalent vaccine.
