Expression, purification and characterization of recombinant murine granulocyte-macrophage colony-stimulating factor and bovine interleukin-2 from yeast.
Gene
; 55(2-3): 287-93, 1987.
Article
em En
| MEDLINE
| ID: mdl-3311885
ABSTRACT
Expression and secretion of two lymphokines, murine granulocyte-macrophage colony-stimulating factor (MuGM-CSF) and bovine interleukin-2 (BoIL-2), to levels of 50-60 mg per liter were achieved by placing these cDNAs in a Saccharomyces cerevisiae expression vector that utilized the yeast alcohol dehydrogenase-2 promoter and alpha-factor leader peptide. These lymphokines were purified to homogeneity by direct application of the crude yeast medium to reversed-phase high-performance liquid chromatography. Despite the fact that both lymphokines contain at least one N-glycosylation site and have identical N-terminal residues (Ala-Pro-Thr), recombinant (R) GM-CSF was found to be heterogeneously glycosylated by yeast while RBoIL-2 was secreted without glycosylation. Additionally, approximately 40% of the RGM-CSF was found to be proteolytically cleaved after the second amino acid residue, while RBoIL-2 was found to be intact.
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Base de dados:
MEDLINE
Assunto principal:
Proteínas Recombinantes de Fusão
/
Proteínas Recombinantes
/
Fatores Estimuladores de Colônias
/
Interleucina-2
Limite:
Animals
Idioma:
En
Ano de publicação:
1987
Tipo de documento:
Article