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A versatile set of vectors for constitutive and regulated gene expression in Pichia pastoris
Author(s) -
Sears Irina B.,
O'Connor James,
Rossanese Olivia W.,
Glick Benjamin S.
Publication year - 1998
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/(sici)1097-0061(19980615)14:8<783::aid-yea272>3.0.co;2-y
Subject(s) - biology , pichia pastoris , multiple cloning site , gene , expression vector , gene expression , genbank , computational biology , vector (molecular biology) , genetics , microbiology and biotechnology , recombinant dna
The budding yeast Pichia pastoris is an attractive system for exploring certain questions in cell biology, but experimental use of this organism has been limited by a lack of convenient expression vectors. Here we describe a set of compact vectors that should allow for the expression of a wide range of endogenous or foreign genes in P. pastoris . A gene of interest is inserted into a modified pUC19 polylinker; targeted integration into the genome then results in stable and uniform expression of this gene. The utility of these vectors was illustrated by expressing the bacterial β‐glucuronidase (GUS) gene. Constitutive GUS expression was obtained with the strong GAP promoter or the moderate YPT1 promoter. The regulatable AOX1 promoter yielded very strong GUS expression in methanol‐grown cells, negligible expression in glucose‐grown cells, and intermediate expression in mannitol‐grown cells. GenBank Accession Numbers are: pIB1, AF027958; pIB2, AF027959; pIB3, AF027960; pIB4, AF027961. © 1998 John Wiley & Sons, Ltd.