title: A constitutive expression system for Pichia pastoris based on the PGK1 promoter.
abstract: OBJECTIVE
  To develop a new vector for constitutive expression in Pichia pastoris based on the endogenous glycolytic PGK1 promoter.
  
  
  RESULTS
  P. pastoris plasmids bearing at least 415 bp of PGK1 promoter sequences can be used to drive plasmid integration by addition at this locus without affecting cell growth. Based on this result, a new P. pastoris integrative vector, pPICK2, was constructed bearing some features that facilitate protein production in this yeast: a ~620 bp PGK1 promoter fragment with three options of restriction sites for plasmid linearization prior to yeast transformation: a codon-optimized α-factor secretion signal, a new polylinker, and the kan marker for vector propagation in bacteria and selection of yeast transformants.
  
  
  CONCLUSIONS
  A new constitutive vector for P. pastoris represents an alternative platform for recombinant protein production and metabolic engineering purposes.
authors.affiliation: Departamento de Biologia Celular, Instituto de Ciências Biológicas, Universidade de Brasília, Brasília, DF, 70901-900, Brazil.
authors.forename: Andrelisse
authors.initials: A
authors.lastname: Arruda
authors.affiliation: Departamento de Biologia Celular, Instituto de Ciências Biológicas, Universidade de Brasília, Brasília, DF, 70901-900, Brazil.
authors.forename: Viviane Castelo Branco
authors.initials: VC
authors.lastname: Reis
authors.affiliation: Departamento de Biologia Celular, Instituto de Ciências Biológicas, Universidade de Brasília, Brasília, DF, 70901-900, Brazil.
authors.forename: Vinícius Daniel Ferreira
authors.initials: VD
authors.lastname: Batista
authors.affiliation: Departamento de Biologia Celular, Instituto de Ciências Biológicas, Universidade de Brasília, Brasília, DF, 70901-900, Brazil.
authors.forename: Bruno Sahim
authors.initials: BS
authors.lastname: Daher
authors.affiliation: Departamento de Biologia Celular, Instituto de Ciências Biológicas, Universidade de Brasília, Brasília, DF, 70901-900, Brazil.
authors.forename: Luiza Cesca
authors.initials: LC
authors.lastname: Piva
authors.affiliation: Departamento de Biologia Celular, Instituto de Ciências Biológicas, Universidade de Brasília, Brasília, DF, 70901-900, Brazil.
authors.forename: Janice Lisboa
authors.initials: JL
authors.lastname: De Marco
authors.affiliation: Departamento de Biologia Celular, Instituto de Ciências Biológicas, Universidade de Brasília, Brasília, DF, 70901-900, Brazil.
authors.forename: Lidia Maria Pepe
authors.initials: LM
authors.lastname: de Moraes
authors.affiliation: Departamento de Biologia Celular, Instituto de Ciências Biológicas, Universidade de Brasília, Brasília, DF, 70901-900, Brazil. ftorres@unb.br.
authors.forename: Fernando Araripe Gonçalves
authors.initials: FA
authors.lastname: Torres
mesh_terms: D015870:Gene Expression; D018390:Gene Targeting / Q000379:methods*; D005822:Genetic Vectors; D005827:Genetics, Microbial / Q000379:methods*; D010735:Phosphoglycerate Kinase / Q000235:genetics*; D010843:Pichia / Q000201:enzymology / Q000235:genetics*; D010957:Plasmids; D011401:Promoter Regions, Genetic
chemical_list: D010735:Phosphoglycerate Kinase
keywords: 3-Phosphoglycerate kinase; Constitutive expression; Pichia pastoris; Recombinant protein production; α-Amylase