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PMID: 3108861 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

Expression of the lacZ gene from two methanol-regulated promoters in Pichia pastoris.

Nucleic acids research ·Vol. 15 ·No. 9 ·1987-05-11 ·Pages 3859-76

Tschopp JF, Brust PF, Cregg JM, Stillman CA, Gingeras TR

Abstract

Two DNA fragments containing putative control regions regulating the expression of the alcohol oxidase (AOX) and dihydroxy-acetone synthase (DAS) genes from the methylotrophic yeast Pichia pastoris were used in the construction of vectors for the expression of the Escherichia coli lacZ gene. These vectors were transformed into P. pastoris host cells and employed in experiments to measure the control mechanisms employed by each promoter in the production of beta-galactosidase fusion products. Results in P. pastoris suggest that the processes used to regulate the expression of these gene fusions involve both repression/derepression and induction mechanisms. Expression of the AOX-lacZ and DAS-lacZ fusions was examined in Saccharomyces cerevisiae as well. Interestingly, beta-galactosidase was expressed in a regulated manner in the heterologous host.

MeSH Terms
Alcohol Oxidoreductases/genetics Aldehyde-Ketone Transferases Bacterial Proteins/biosynthesis,genetics Escherichia coli/genetics Fungal Proteins/genetics Galactosidases/biosynthesis Gene Expression Regulation Genes, Fungal Methanol/pharmacology Pichia/genetics Promoter Regions, Genetic/drug effects Recombinant Fusion Proteins/biosynthesis,genetics Recombinant Proteins/biosynthesis Saccharomyces cerevisiae/genetics Saccharomycetales/genetics Transferases/genetics beta-Galactosidase/biosynthesis,genetics
Chemicals
Bacterial Proteins Fungal Proteins Recombinant Fusion Proteins Recombinant Proteins Alcohol Oxidoreductases alcohol oxidase Transferases Aldehyde-Ketone Transferases formaldehyde transketolase Galactosidases beta-Galactosidase Methanol
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Tschopp J F
Brust P F
Cregg J M
Stillman C A
Gingeras T R
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31 references, click to expand
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/15.9.3859
Published
1987-05-11
Pages
3859-76
Language
English
Region
England
NLM ID
0411011
PMCID
PMC340787
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/15.9.3859 PMC Full Text View journal details
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