[Expression, purification and characterization of the recombinant anthrax protective antigen].
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.
Xu
Jun-Jie
JJ
Institute of Microbiology and Epidemiology, Beijing 100071, China.
Dong
Da-Yong
DY
Song
Xiao-Hong
XH
Ge
Meng
M
Li
Guan-Lin
GL
Fu
Ling
L
Zhuang
Han-Lan
HL
Chen
Wei
W
D000595:Amino Acid Sequence; D000818:Animals; D022122:Anthrax Vaccines / Q000276:immunology*; D000942:Antigens, Bacterial / Q000737:chemistry / Q000235:genetics* / Q000276:immunology / Q000633:toxicity; D001427:Bacterial Toxins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology / Q000633:toxicity; D001483:Base Sequence; D051379:Mice; D008969:Molecular Sequence Data; D010957:Plasmids; D011817:Rabbits; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000276:immunology; D014614:Vaccines, Synthetic / Q000276:immunology
D022122:Anthrax Vaccines; D000942:Antigens, Bacterial; D001427:Bacterial Toxins; D011994:Recombinant Proteins; D014614:Vaccines, Synthetic; C030325:anthrax toxin