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Cloning, sequencing and expression of the transferrin-binding protein 2 gene from Actinobacillus pleuropneumoniae

Published online by Cambridge University Press:  12 February 2007

Wang Fang  and
He Kong-Wang
Wang Fang
Affiliation:
Key Laboratory of Animal and Poultry Diseases Diagnostic, Ministry of Agriculture; Institute of Veterinary Medicine of Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China
He Kong-Wang*
Affiliation:
Key Laboratory of Animal and Poultry Diseases Diagnostic, Ministry of Agriculture; Institute of Veterinary Medicine of Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China
*
*Corresponding author: Email [email protected]
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Abstract

To amplify the transferrin-binding protein 2 (Tbp2) gene from Actinobacillus pleuropneumoniae (App) by polymerase chain reaction (PCR), a pair of primers were designed according to sequence Z46774 of A. pleuropneumoniae serotype 5. The amplified DNA fragment (1642 bp) was cloned into pMD18-T and sequenced. The sequencing result showed that the homology was 99.8% compared with the reference sequence. The target fragment of the positive clones was inserted into pET-32a and transformed in Escherichia coli BL21 (DE3). After its induction, the fragment was expressed in E. coli. The Western blotting results were positive.

Keywords

Actinobacillus pleuropneumoniaecloningexpressionsequencingtransferrin-binding protein 2
Type
Research Article
Information
Chinese Journal of Agricultural Biotechnology , Volume 2 , Issue 3 , December 2005 , pp. 155 - 159
Copyright
Copyright © China Agricultural University and Cambridge University Press 2005

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References

Ausubel, FM, Brent, R and Kingston, RE (2001) Short Protocols in Molecular Biology. Beijing: Science Press (in Chinese).CrossRefGoogle Scholar
Baltes, N, Hennig-Pauka, I and Gerlach, GF (2002) Both transferrin binding proteins are virulence factors in Actinobacillus pleuropneumoniae serotype 7 infection. FEMS Microbiology Letters 209: 283–287.CrossRefGoogle ScholarPubMed
Blackall, PJ, Klaasen, HL, van den Bosch, H, Kuhnert, P and Frey, J (2002) Proposal of a new serovar of Actinobacillus pleuropneumoniae: serovar 15. Veterinary Microbiology 84: 47–52.CrossRefGoogle ScholarPubMed
Bosse, JT, Janson, H, Sheehan, BJ et al. , (2002) Actinobacillus pleuropneumoniae: pathobiology and pathogenesis of infection. Microbes and Infection 4: 225–235.Google Scholar
Ferreiros, CM, Ferron, L and Criado, MT (1994) In vivo human immune response to transferrin-binding protein 2 and other iron-regulated proteins of Neisseria meningitidis. FEMS Immunology and Medicine Microbiology 8: 63–68.Google Scholar
Guerinot, ML (1994) Microbial iron transport. Annual Reviews of Microbiology 48: 743–772.CrossRefGoogle ScholarPubMed
Jarma, E and Regassa, LB (2004) Growth phase mediated regulation of the Actinobacillus pleuropneumoniae ApxI and ApxII toxins. Microbial Pathogenesis 36: 197–203.Google Scholar
Lissolo, L, Maitre-Wilmotte, G, Dumas, P, Mignon, M, Danve, B and Quentin-Millet, MJ (1995) Evaluation of transferrin-binding protein 2 within the transferrin-binding protein complex as a potential antigen for future meningococcal vaccines. Infection and Immunity 63: 884–890.CrossRefGoogle ScholarPubMed
Litt, DJ, Palmer, HM and Borriello, SP (2000) Neisseria meningitidis expressing transferrin binding proteins of Actinobacillus pleuropneumoniae can utilize porcine transferrin for growth. Infection and Immunity 68: 550–557.CrossRefGoogle ScholarPubMed
Myers, LE, Yang, YP, Du, RP et al. , (1998) The transferrin binding protein B of Moraxella catarrhalis elicits bactericidal antibodies and is a potential vaccine antigen. Infection and Immunity 66: 4183–4192.Google Scholar
Pattison, IH, Howell, DG and Elliott, J (1957) A Haemophilus-like organism isolated from pig lung and the associated pneumonic lesions. Journal of Comparative Pathology 67: 320–329.CrossRefGoogle ScholarPubMed
Sambrook, J and Russell, DW (2002) Molecular Cloning: A Laboratory Manual, 3rd ed. Beijing: Science Press (in Chinese).Google Scholar
Van Overbeke, I, Chiers, K, Ducatelle, R and Haesebrouck, F (2001) Effect of endobronchial challenge with Actinobacillus pleuropneumoniae serotype 9 of pigs vaccinated with a vaccine containing Apx toxins and transferrin-binding proteins. Journal of Veterinary Medicine, Series B 48: 15–20.Google Scholar