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- W2393023976 abstract "Objective To clone porcine interferon α(IFNα)gene and express in Pichia pastoris. Methods Lymphocytes were isolated from porcine peripheral blood and induced by phytohemagglutinin(PHA), from which total RNA was extracted with Trizol reagent for amplification of porcine IFNα gene by RT-PCR. The amplified gene was cloned into vector pPIC9K, and the constructed recombinant plasmid pPIC9K-IFNα was identified by PCR, restriction analysis and sequencing, then transformed to P. pastoris GS115 by electrotransformation. The recombinants were screened for expression under induction of methanol. The expressed product was identified by SDS-PAGE and Western blot. Results The porcine IFNα gene at a length of 521 bp was amplified by RT-PCR. PCR, restriction analysis and sequencing proved that recombinant plasmid pPIC9K-IFNα was constructed correctly. The expressed protein showed a relative molecular mass of about 19 000, and that in soluble form contained about 13% of total somatic protein. Specific immune reaction of the expressed protein with the corresponding antibody was observed. Conclusion Porcine IFNα gene was successfully cloned and expressed in P. pastoris, which laid a foundation of further study on biological characters of porcine IFNα." @default.
- W2393023976 created "2016-06-24" @default.
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- W2393023976 date "2010-01-01" @default.
- W2393023976 modified "2023-09-24" @default.
- W2393023976 title "Cloning and Eukaryotic Expression of Porcine Interferon α Gene" @default.
- W2393023976 hasPublicationYear "2010" @default.
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