Authors
Chaves, Julie; Wilton, Rosemarie; Gao, Yuqian; Munoz Munoz, Nathalie; Burnet, Meagan; Schmitz, Zachary; Rowan, John; Burdick, Leah; Elmore, Joshua; Guss, Adam; Close, Dan; Magnuson, Jon; Burnum-Johnson, kristin; Michener, Joshua
Abstract
The development ofPseudomonasstrains for industrial production of fuels and chemicals will require the integration of heterologous genes and pathways into thechromosome. Finding the most appropriate integration site to maximize strain performance is an essential part of the strain design process. We characterized sevenchromosomal loci inPseudomonas putidaKT2440 for integration of afluorescent protein expression construct. Insertion infive of the loci did not affect growth rate, butfluorescence varied by up to 27-fold. Three sites displaying a diversity of phenotypes with thefluorescent reporter were also chosen for the integration of a geneencoding a muconate importer. Depending on the integration locus, expression of the importer varied by approximately 3-fold and produced significant phenotypicdifferences. This work demonstrates the impact of the integration location on host viability, gene expression, and overall strain performance.