A versatile set of vectors for constitutive and regulated gene expression in Pichia pastoris.
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 beta-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, AF0279959; pIB3, AF027960; pIB4, AF027961.
Sears
I B
IB
Department of Molecular Genetics and Cell Biology, University of Chicago, IL 60637, USA.
O'Connor
J
J
Rossanese
O W
OW
Glick
B S
BS
D000429:Alcohol Oxidoreductases / Q000235:genetics; D000619:Aminohydrolases; D004790:Enzyme Induction; D005656:Fungal Proteins / Q000235:genetics; D020558:GTP Phosphohydrolases / Q000235:genetics; D019204:GTP-Binding Proteins / Q000235:genetics; D015966:Gene Expression Regulation, Fungal; D018390:Gene Targeting; D017930:Genes, Reporter / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics*; D005947:Glucose; D005966:Glucuronidase / Q000378:metabolism; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics; D008353:Mannitol; D000432:Methanol; D008969:Molecular Sequence Data; D010843:Pichia / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics; D011755:Pyrophosphatases; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors / Q000235:genetics; D020691:rab GTP-Binding Proteins
D005656:Fungal Proteins; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D008353:Mannitol; D000429:Alcohol Oxidoreductases; C092116:HIS4 protein, S cerevisiae; C019643:alcohol oxidase; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D005966:Glucuronidase; D000619:Aminohydrolases; D020558:GTP Phosphohydrolases; D019204:GTP-Binding Proteins; D011755:Pyrophosphatases; C060755:YPT1 protein, S cerevisiae; D020691:rab GTP-Binding Proteins; D005947:Glucose; D000432:Methanol