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A versatile set of vectors for constitutive and regulated gene expression inPichia pastoris

✍ Scribed by Sears, Irina B.; O'Connor, James; Rossanese, Olivia W.; Glick, Benjamin S.

Publisher: John Wiley and Sons
Year: 1998
Tongue: English
Weight: 137 KB
Volume: 14
Category: Article
ISSN: 0749-503X
DOI: 10.1002/(sici)1097-0061(19980615)14:8<783::aid-yea272>3.0.co;2-y

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✦ Synopsis

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 -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.

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