Title: Construction and expression of gene encoding interleukin-2/human serum album fusion protein in Pichia pastria
Abstract:Aim:To construct the recombinant plasmid pPIH encoding IL-2-human serum album(HSA) fusion protein, express it in Pichia pastria, and to obtain the IL-2-HSA with IL-2 bioactivity. Methods: The cDNA enc...Aim:To construct the recombinant plasmid pPIH encoding IL-2-human serum album(HSA) fusion protein, express it in Pichia pastria, and to obtain the IL-2-HSA with IL-2 bioactivity. Methods: The cDNA encoding human IL-2 was obtained by RT-PCR from human peripheral blood mononuclear cells. The two cDNA fragments encoding human IL-2 and HSA respectively were spliced by overlap PCR extension, and the fusion gene was then cloned into the expression vector pPIC9K to obtain the recombinant plasmid pPIH. The recombinant P. pastria determined was obtained by electrotransformation of pPIH into P. pastria KM71. The bioactivity of the supernatant containing the fusion protein IL-2-HSA was assayed by CTLL-2/MTT method. Results: The recombinant plasmid pPIH was successfully constructed, and the fusion protein IL-2-HSA was well expressed from pPIH with methanol induction at 30℃ for 3 d. It was found that the bioactivity of the fermentation supernatant was about 100 U/mL. Conclusion: The recombinant fusion protein of IL-2-HSA with IL-2 bioactivity was successfully obtained. The proposed method for the recombinant fusion protein of IL-2-HSA has not been reported.Read More
Publication Year: 2008
Publication Date: 2008-01-01
Language: en
Type: article
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Cited By Count: 1
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