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

Yeast. 1998 Jun 15;14(8):783-90. doi: 10.1002/(SICI)1097-0061(19980615)14:8<783::AID-YEA272>3.0.CO;2-Y.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Alcohol Oxidoreductases / genetics
  • Aminohydrolases
  • Enzyme Induction
  • Fungal Proteins / genetics
  • GTP Phosphohydrolases / genetics
  • GTP-Binding Proteins / genetics
  • Gene Expression Regulation, Fungal
  • Gene Targeting
  • Genes, Reporter / genetics
  • Genetic Vectors / genetics*
  • Glucose
  • Glucuronidase / metabolism
  • Glyceraldehyde-3-Phosphate Dehydrogenases / genetics
  • Mannitol
  • Methanol
  • Molecular Sequence Data
  • Pichia / genetics*
  • Promoter Regions, Genetic / genetics
  • Pyrophosphatases
  • Saccharomyces cerevisiae Proteins*
  • Transcription Factors / genetics
  • rab GTP-Binding Proteins*

Substances

  • Fungal Proteins
  • Saccharomyces cerevisiae Proteins
  • Transcription Factors
  • Mannitol
  • Alcohol Oxidoreductases
  • HIS4 protein, S cerevisiae
  • alcohol oxidase
  • Glyceraldehyde-3-Phosphate Dehydrogenases
  • Glucuronidase
  • Aminohydrolases
  • GTP Phosphohydrolases
  • GTP-Binding Proteins
  • Pyrophosphatases
  • YPT1 protein, S cerevisiae
  • rab GTP-Binding Proteins
  • Glucose
  • Methanol

Associated data

  • GENBANK/AF027958
  • GENBANK/AF027959
  • GENBANK/AF027960
  • GENBANK/AF027961