Efficient intra- and extracellular production of human beta-1,4-galactosyltransferase in Saccharomyces cerevisiae is mediated by yeast secretion leaders.
Gene
; 145(2): 299-303, 1994 Aug 05.
Article
em En
| MEDLINE
| ID: mdl-8056347
ABSTRACT
We have compared the function of homologous and heterologous secretion leaders to mediate production of human beta-1,4-galactosyltransferase (Gal-Tf) in the yeast Saccharomyces cerevisiae. Although all genes encoding leader/Gal-Tf fusions were transcribed by strong yeast promoters, only low production levels were obtained for full-length Gal-Tf containing the human membrane-anchor region. In contrast, a gene fusion encoding the membrane-anchor region of the yeast alpha-1,2-mannosyltransferase (Mnt1) fused to soluble Gal-Tf yielded high mRNA and intracellular protein levels. Gal-Tf could also be produced extracellularly using a fusion of the pre-pro region of the yeast Mf alpha 1 precursor (MF alpha 1) to soluble Gal-Tf; a fusion containing only the pre-region of Mf alpha 1 was synthesized intracellularly, but did not lead to Gal-Tf activity in the culture medium. All yeast-produced Gal-Tf proteins were enzymatically active. These results demonstrate that yeast secretion leaders are advantageous to achieve efficient production of active Gal-Tf in S. cerevisiae.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Sinais Direcionadores de Proteínas
/
N-Acetil-Lactosamina Sintase
/
Proteínas de Saccharomyces cerevisiae
Limite:
Humans
Idioma:
En
Revista:
Gene
Ano de publicação:
1994
Tipo de documento:
Article
País de afiliação:
Alemanha