Development and characterization of a vector set with regulated promoters for systematic metabolic engineering in Saccharomyces cerevisiae

作者：Shen Michael W Y; Fang Fang; Sandmeyer Suzanne; Da Silva Nancy A^*

来源：Yeast, 2012, 29(12): 495-503.

DOI：10.1002/yea.2930

• Saccharomyces cerevisiae
• vectors
• integration
• inducible yeast promoters
• Cre recombinase
• metabolic engineering

A set of vectors was constructed that enable combined and systematic testing of metabolic pathway genes in Saccharomyces cerevisiae. The vectors are available as CEN/ARS and 2 mu based plasmids with a choice of three inducible promoters, PGAL1, PCUP1 and PADH2. These features offer control over the initiation and level of gene expression. In addition, the vectors can be used as templates to generate PCR fragments for targeted chromosomal integration of gene expression cassettes. Selection markers are flanked by loxP elements to allow efficient CreA-mediated marker removal and recycling after genomic integration. For each promoter, expression of a bacterial lacZ reporter gene was characterized from plasmid-based and integrated chromosomal cassettes, and compared to that of the glycolytic PPGK1 promoter. Plasmid stabilities were also determined. The promoters showed distinct activity profiles useful for modulating expression of metabolic pathway genes. This series of plasmids with inducible promoters extends our previous vector set carrying the constitutive promoters PPGK1, PTEF1 and PHXT7-391.

• 出版日期2012-12