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Expressing, Purification and Identification of IFN-tau Gene in the E.coli |
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Abstract Interferon-tau (IFN-tau) is a type I IFN originally discovered for its role as a pregnancy recognition hormone in ruminant animals such as sheep and cows. IFN-tau possesses all of the biological properties ascribed to the other type I IFNs including antiviral, antiproliferative and immunomodulatory activities. In order to identify the function of IFN-tau clearly, the coding sequence of IFN-tau was amplified by PCR from IFN-tau cDNA, this fragment digested by EcoR I and BamH I was inserted into pBV220 by T4 ligase and transformed into E.coli BL21 then the recombinant plasmid named pBV220/IFN-tau was constructed successfully. IFN-tau was expressed distinctly after inducing by the temperature. The expression production was purified on S-100 High Resolution. The product was identified as IFN-tau by amino acid sequence analysis. The specific activity of IFN-tau was about 2.09×106IU/ml assayed by the standard antiviral activity test on WISH cells challenged with VSV virus.
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Received: 16 August 2006
Published: 25 March 2007
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