Korean Journal of Veterinary Research (대한수의학회지)

The Korean Society of Veterinary Science (대한수의학회)
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Mass expression of Apx I and Apx II of Actinobacillus pleuropneumoniae in Escherichia coli

대장균에서 흉막폐렴균 독소 Apx I과 Apx II의 대량발현

  • Kim, Tae-Jung (College of Veterinary Medicine, Chonnam National University) ;
  • Lee, Bong-Joo (College of Veterinary Medicine, Chonnam National University) ;
  • Lee, Jae-Il (College of Veterinary Medicine, Chonnam National University)
  • Accepted : 2005.05.20
  • Published : 2005.06.30
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Abstract

Actinobacillus pleuropneumoniae is the causative agent of a porcine contagious pleuropneumonia. Among several virulence factors including exotoxin (Apx toxins), LPS, transferrin-binding proteins, OMPs, and some proteases, Apx toxins have been major targets for the protection study. In this study, cloning and expression of A. pleuropneumoniae Apx I and Apx II toxin, which are produced by all highly virulent strains, were performed by Escherichia coli expression system. Genes coding Apx I and II toxin were amplified from the A. pleuropneumoniae serotype 5 genomic DNA using polymerase chain reaction and cloned to a prokaryotic expression vector, pRSET. Expression of the Apx I and Apx II coding sequences in E. coli resulted in the formation of insoluble inclusion bodies purified according to a denaturing purification protocol, which employs the use of guanidium. Recombinant proteins were purified using $Ni^{2+}$-charged resin affinity purification. This expression and purification system made it possible to produce Apx I and Apx II in large amounts for further immunologic studies.

Keywords

References

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