[Expression and characterization of the recombination anthrax lethal factor].
The gene encoding anthrax lehtal factor (LF) was cloned into a secretory expression plasmid and then expressed in periplasmic space of E. coli. The recombinant LF (rLF) expressed was about 4% of the total proteins in E. coli. About 3 mg electrophoresis purity rLF could be obtained after the purification of 1 liter culure using ion exchange chromatography and gel filtration. The result of sequencing assay showed that the N-terminal amino acid sequence of rLF was identical to the N-terminal sequence of natural LF. In vitro toxicity analysis also shows that rLF has an excellent biological activity. The successful expression of rLF has placed a solid foundation for the research on toxicity mechanism of LF, developing new anthrax vaccines, and screening for inhibitors against anthrax toxin.
Guo
Qiang
Q
Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071, China. timeguo@sohu.com
Xu
Jun-Jie
JJ
Dong
Da-Yong
DY
Fu
Ling
L
Chen
Wei
W
D000818:Animals; D000942:Antigens, Bacterial / Q000235:genetics* / Q000302:isolation & purification / Q000633:toxicity; D001427:Bacterial Toxins / Q000235:genetics* / Q000302:isolation & purification / Q000633:toxicity; D004926:Escherichia coli / Q000235:genetics; D051379:Mice; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification
D000942:Antigens, Bacterial; D001427:Bacterial Toxins; D011994:Recombinant Proteins; C030325:anthrax toxin