Hostname: page-component-586b7cd67f-t8hqh Total loading time: 0 Render date: 2024-11-24T08:41:38.512Z Has data issue: false hasContentIssue false
  • English
  • Français

Water buffalo (Bubalus bubalus) interferon-α genes: cloning, expression and antiviral activities

Published online by Cambridge University Press:  12 February 2007

Shi Xi-Ju ,
Xia Chun  and
Wang Ming
Shi Xi-Ju
Affiliation:
College of Veterinary Medicine, China Agricultural University, Beijing 100094, China
Xia Chun
Affiliation:
College of Veterinary Medicine, China Agricultural University, Beijing 100094, China
Get access Rights & Permissions [Opens in a new window]

Abstract

Interferon (IFN)-α genes were cloned from genomic DNA of Fuan and Fuzhong water buffaloes by PCR, and the PCR products were inserted into a pQE30 vector to construct recombinant expression plasmids. Sequence analysis showed that both clones were composed of 498 nucleotides, encoding a mature polypeptide with 166 amino acids (aa). They were defined as two new subtypes, with 91.6–94.2% identity at the amino acid level by comparison with eight previously published bovine IFN-α subtypes. Results of SDS-PAGE and Western blotting showed that each of the recombinant proteins was expressed in inclusion bodies in Escherichia coli with molecular weight of 20 kDa and the recombinant proteins were 25% of the whole proteins. Inclusion bodies were denatured and renatured with urea and the antiviral activities of the recombinant buffalo IFN-α (rBuIFN-α) were 105 U/mg and 106 U/mg in CEF/VSV and MDBK/VSV cell lines, respectively. Additionally, rBuIFN-α had good effects against challenge by infectious bovine rhinotracheitis virus. The rBuIFN-α are potential biological agents for the prevention and treatment of various kinds of bovine viral disease.

Keywords

Fuan and Fuzhong water buffalointerferon-α geneantiviral activities
Type
Research Article
Information
Chinese Journal of Agricultural Biotechnology , Volume 2 , Issue 1 , April 2005 , pp. 45 - 52
Copyright
Copyright © China Agricultural University and Cambridge University Press 2005

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Baruch, V, Sara, C, Haim, G, Leitner, M and Shafferman, A (1985) Bovine interferon α genes. Journal of Biological Chemistry 260: 5498–5504.Google Scholar
Capon, DJ, Shephard, HM and Goeddel, DV (1985) Two distinct families of human and bovine interferon-α genes are coordinately expressed and encode functional polypeptides. Molecular and Cellular Biology 5: 768–779.Google ScholarPubMed
Chaplin, PJ, Entrican, G, Gelder, KI and Collins, RA (1996) Cloning and biologic activities of a bovine interferon-α isolated from the epithelium of a rotavirus-infected calf. Journal of Interferon and Cytokine Research 16: 25–30.CrossRefGoogle ScholarPubMed
Chaplin, PJ, Parsons, KR and Collins, RA (1996) The cloning of cattle interferon-A subtypes isolated from the gut epithelium of rotavirus-infected calves. Immunogenetics 44: 143–145.Google Scholar
Charleston, B, Fray, MD, Baigent, S, Carr, BV and Morrison, WI (2001) Establishment of persistent infection with non-cytopathic bovine viral diarrhoea virus in cattle is associated with a failure to induce type I interferon. Journal of General Virology 82: 1893–1897.Google Scholar
Chinsangaram, J, Piccone, ME and Grubman, MJ (1999) Ability of foot-and-mouth disease virus to form plaques in cell culture is associated with suppression of alpha/beta interferon. Journal of Virology 73: 9891–9898.CrossRefGoogle ScholarPubMed
Esparza, I, Gonzalez, JC and Vinuela, E (1988) Effect of interferon-α, interferon-γ and tumour necrosis factor on African swine fever virus replication in porcine monocytes and macrophages. Journal of General Virology 69: 2973–2980.CrossRefGoogle ScholarPubMed
Jacobsen, KL, Rockwood, GA, Abolhassani, M et al. , (1988) Kinetics of large-scale production of bovine leukocyte interferon, using three viral inducers. American Journal of Veterinary Research 49: 1441–1446.Google ScholarPubMed
Perler, L, Schweizer, M, Jungi, T and Perterhas, E (2000) Bovine viral diarrhea virus and bovine herpesvirus-1 prime uninfected macrophages for lipoplysaccharide-triggered apoptosis by interferon-dependent and -independent pathways. Journal of Gene Virology 81: 881–887.Google Scholar
Roberts, RM, Liu, LM, Guo, QT, Leaman, D and Bixby, J (1998). The evolution of the type I interferons. Journal of Interferon and Cytokine Research 18: 805–816.CrossRefGoogle ScholarPubMed
Sambrook, J, Fritsch, EF and Maniatis, T (2001) Molecular Cloning: A Laboratory Manual, 3rd ed. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press.Google Scholar
Wan, JQ, Wu, WX and Xia, C (2002) Expression of porcine gamma-IFN in Pichia pastoris and its effect against PRRSV. Chinese Journal of Biotechnology 18: 683–686 (in Chinese with English abstract).Google Scholar
Wang, M, Wu, ZG and Xia, C (2000) Cloning, sequence analysis and expression in E. coli of IFN-alpha gene of AA chickens. Journal of Agricultural Biotechnology 8: 377–381 (in Chinese with English abstract).Google Scholar
Wu, ZG, Xia, C and Wang, M (2002) Studies on expression of chicken IFN-γ cDNA in QIAexpress expression system. Journal of China Agricultural University 7: 89–93 (in Chinese with English abstract).Google Scholar