Renaturation and purification of ApxII toxin of Actinobacillus pleuropneumoniae.
ApxII toxin is the only Apx toxin that is produced by Actinobacillus pleuropneumoniae serotype 7. In order to determine whether the recombinant ApxII that derived from Escherichia coli (E. coli) expression is faithful to the natural ApxII so that can be used as additional component in vaccine preparation, the structure gene apxIIA of ApxII toxin was expressed in E. coli with prokaryotic expression vector pGEX-6p-1 (formed pGEX-6p-A). pGZRS-C which is A. pleuropneumoniae-E. coli shuttle vector pGZRS-38 expressing the post-transcriptional activation gene apxII C was co-expressed with pGEX-6p-A. The expression product of rApxII A formed inclusion. The inclusion protein was oxidized, refolded and restored hemolytic activity after denaturation, renaturation and purification. The result indicated that E. coli expressed recombinant ApxII toxin has good fidelity, which makes it possible to produce this valuable antigen for vaccine preparation or diagnosis.
Wang
Chunlai
C
Division of Bacterial Diseases, National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, PR China. chunlaiwang555@hotmail.com
Liu
Siguo
S
Peng
Yonggang
Y
Shao
Meili
M
Wang
Yong
Y
Gong
Qiang
Q
Chang
Yuehong
Y
Liu
Jiandong
J
Liu
Huifang
H
Liu
Di
D
Kong
Xiangang
X
D016977:Actinobacillus pleuropneumoniae / Q000737:chemistry*; D001426:Bacterial Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D006460:Hemolysin Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D006461:Hemolysis
C085361:ApxII toxin, bacteria; D001426:Bacterial Proteins; D006460:Hemolysin Proteins