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Expression of Recombinant Proteins in the Methylotrophic Yeast Pichia pastoris
Published on: February 25, 2010
Production of recombinant human placental variant growth hormone in Pichia pastoris
José Prisco Palma-Nicolás1, Jorge A Ascacio-Martínez, Agnès Revol
1Departamento de Bioquímica, Laboratorio de Biotecnología, Facultad de Medicina de la Universidad Autónoma de Nuevo León, Ave. Madero y Dr. Eduardo Aguirre Pequeño, Col. Mitras Centro, 64460 Monterrey, N.L., México.
Abstract:
cDNA encoding mature human placental variant growth hormone (HGH-V) was synthesized by retro-transcription polymerase chain reaction (RT-PCR) from total RNA recovered from human term-placenta and cloned in pBluescript plasmid (pBS) in Escherichia coli. cDNA was subcloned into pPIC9, fusing it to the flanking regulatory sequences of the Pichia pastoris alcohol oxidase 1 gene (AOX1) and finally introduced into the genome of this yeast by homologous recombination. The resulting new recombinant strain produced and secreted, towards the culture medium, mature HGH-V, whose activity was demonstrated in cell culture by the Nb2 proliferation assay.
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