Expression, purification and characterization of the recombinant anthrax protective antigen.
- Author:
Jun-Jie XU
1
;
Da-Yong DONG
;
Xiao-Hong SONG
;
Meng GE
;
Guan-Lin LI
;
Ling FU
;
Han-Lan ZHUANG
;
Wei CHEN
Author Information
1. Institute of Microbiology and Epidemiology, Beijing 100071, China.
- Publication Type:Journal Article
- MeSH:
Amino Acid Sequence;
Animals;
Anthrax Vaccines;
immunology;
Antigens, Bacterial;
chemistry;
genetics;
immunology;
toxicity;
Bacterial Toxins;
chemistry;
genetics;
immunology;
toxicity;
Base Sequence;
Mice;
Molecular Sequence Data;
Plasmids;
Rabbits;
Recombinant Proteins;
biosynthesis;
immunology;
Vaccines, Synthetic;
immunology
- From:
Chinese Journal of Biotechnology
2004;20(5):652-655
- CountryChina
- Language:Chinese
-
Abstract:
An expression plasmid carrying anthrax protective antigen (PA) gene was constructed, which has an OmpA signal sequence attached to the 5' end of PA gene. The plasmid was transformed into E. coli and induced to express recombinant PA (rPA) . The recombinant protein, about 10% of the total bacterial protein in volume, was secreted to the periplasmic space of the cell. After a purification procedure including ion-exchange, hydrophobic interaction chromatography, and gel filtration, about 15 mg of 95 % pure rPA was obtained from 1-liter culture. The bioactivity of rPA was proved by in vitro cytotoxicity assay. The polyclonal antiserum from rabbits immunized with rPA could inhibit the action of anthrax lethal toxin in vitro, which suggests that antibodies against rPA can provide high passive protection against anthrax. The results reported here may be helpful to develop a safe and efficacious recombinant PA vaccine against anthrax.
