A constitutive expression system for Pichia pastoris based on the PGK1 promoter

作者：Arruda Andrelisse; Branco Reis Viviane Castelo; Ferreira Batista Vinicius Daniel; Daher Bruno Sahim; Piva Luiza Cesca; De Marco Janice Li**oa; Pepe de Moraes Lidia Maria; Goncalves Torres Fernando Araripe

来源：Biotechnology Letters, 2016, 38(3): 509-517.

DOI：10.1007/s10529-015-2002-2

• alpha-Amylase
• Constitutive expression
• 3-Phosphoglycerate kinase
• Pichia pastoris
• Recombinant protein production

Objectives 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 similar to 620 bp PGK1 promoter fragment with three options of restriction sites for plasmid linearization prior to yeast transformation: a codon-optimized alpha-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.

• 出版日期2016-3