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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
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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

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