Heterologous expression of human granulocyte-colony stimulating factor in Pichia pastoris

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. © 2008 Asian Network for Scientific Information.