title: Evaluation of promoters and ribosome binding sites for biotechnological applications in the unicellular cyanobacterium Synechocystis sp. PCC 6803.
abstract: For effective metabolic engineering, a toolbox of genetic components that enables predictable control of gene expression is needed. Here we present a systematic study of promoters and ribosome binding sites in the unicellular cyanobacterium Synechocystis sp. PCC 6803. A set of metal ion inducible promoters from Synechocystis were compared to commonly used constitutive promoters, by measuring fluorescence of a reporter protein in a standardized setting to allow for accurate comparisons of promoter activity. The most versatile and useful promoter was found to be PnrsB, which from a relatively silent expression could be induced almost 40-fold, nearly up to the activity of the strong psbA2 promoter. By varying the concentrations of the two metal ion inducers Ni2+and Co2+, expression from the promoter was highly tunable, results that were reproduced with PnrsB driving ethanol production. The activities of several ribosomal binding sites were also measured, and tested in parallel in Synechocystis and Escherichia coli. The results of the study add useful information to the Synechocystis genetic toolbox for biotechnological applications.
authors.affiliation: Department of Chemistry - Ångström, Uppsala University, Box 523, SE-751 20 Uppsala, Sweden.
authors.forename: Elias
authors.initials: E
authors.lastname: Englund
authors.affiliation: Department of Chemistry - Ångström, Uppsala University, Box 523, SE-751 20 Uppsala, Sweden.
authors.forename: Feiyan
authors.initials: F
authors.lastname: Liang
authors.affiliation: Department of Chemistry - Ångström, Uppsala University, Box 523, SE-751 20 Uppsala, Sweden.
authors.forename: Pia
authors.initials: P
authors.lastname: Lindberg
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics; D001665:Binding Sites; D001709:Biotechnology; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D000431:Ethanol / Q000378:metabolism; D005798:Genes, Bacterial; D008027:Light; D008164:Luminescent Proteins / Q000235:genetics; D011401:Promoter Regions, Genetic; D012270:Ribosomes / Q000378:metabolism*; D046939:Synechocystis / Q000235:genetics / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D008164:Luminescent Proteins; C054562:yellow fluorescent protein, Bacteria; D000431:Ethanol