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Biotechnology
Year: 2008  |  Volume: 7  |  Issue: 3  |  Page No.: 569 - 573

Heterologous Expression of Human Granulocyte-Colony Stimulating Factor in Pichia pastoris

Alireza Saeedinia, Mehdi Shamsara, Ali Bahrami, Mehdi Zeinoddini, Mohammad Ali Naseeri-Khalili, Reza Mohammadi, Narges Malek Sabet and Hosseinali Sami    

Abstract: Human  granulocyte-colony  stimulating  factor (hG-CSF) was expression in a eukaryotic system, Pichia pastoris. For this, hG-CSF cDNA was amplified using reverse transcription-polymerase chain reaction (RT-PCR) and inserted into pPIC9 vector. The recombinant plasmid was electroporated into the P. pastoris. The recombinant hG-CSF was successfully expressed by in this system. The protein of approximately 18 kDa was secreted into the culture supernatant when induced with methanol. Western blot showed that the recombinant hG-CSF expressed in P. pastoris had a high specificity to monoclonal antibody against hG-CSF. The expression of hG-CSF was determined 3.1 mg L-1 in shake flask culture by ELISA method.

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