Production of recombinant human placental variant growth hormone in Pichia pastoris.
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.
Palma-Nicolás
José Prisco
JP
Departamento 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.
Ascacio-Martínez
Jorge A
JA
Revol
Agnès
A
Barrera-Saldaña
Hugo A
HA
D001681:Biological Assay; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D005260:Female; D013006:Growth Hormone / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology; D006801:Humans; D010843:Pichia / Q000235:genetics / Q000378:metabolism*; D010926:Placental Hormones / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000493:pharmacokinetics
C064980:GH2 protein, human; D010926:Placental Hormones; D011994:Recombinant Proteins; D013006:Growth Hormone