title: Synthetic biology toolbox for controlling gene expression in the cyanobacterium Synechococcus sp. strain PCC 7002.
abstract: The application of synthetic biology requires characterized tools to precisely control gene expression. This toolbox of genetic parts previously did not exist for the industrially promising cyanobacterium, Synechococcus sp. strain PCC 7002. To address this gap, two orthogonal constitutive promoter libraries, one based on a cyanobacterial promoter and the other ported from Escherichia coli, were built and tested in PCC 7002. The libraries demonstrated 3 and 2.5 log dynamic ranges, respectively, but correlated poorly with E. coli expression levels. These promoter libraries were then combined to create and optimize a series of IPTG inducible cassettes. The resultant induction system had a 48-fold dynamic range and was shown to out-perform Ptrc constructs. Finally, a RBS library was designed and tested in PCC 7002. The presented synthetic biology toolbox will enable accelerated engineering of PCC 7002.
authors.affiliation: †Department of Chemical and Biological Engineering and ‡Microbiology Doctoral Training Program, University of Wisconsin-Madison, Madison, Wisconsin 53706, United States.
authors.forename: Andrew L
authors.initials: AL
authors.lastname: Markley
authors.affiliation: †Department of Chemical and Biological Engineering and ‡Microbiology Doctoral Training Program, University of Wisconsin-Madison, Madison, Wisconsin 53706, United States.
authors.forename: Matthew B
authors.initials: MB
authors.lastname: Begemann
authors.affiliation: †Department of Chemical and Biological Engineering and ‡Microbiology Doctoral Training Program, University of Wisconsin-Madison, Madison, Wisconsin 53706, United States.
authors.forename: Ryan E
authors.initials: RE
authors.lastname: Clarke
authors.affiliation: †Department of Chemical and Biological Engineering and ‡Microbiology Doctoral Training Program, University of Wisconsin-Madison, Madison, Wisconsin 53706, United States.
authors.forename: Gina C
authors.initials: GC
authors.lastname: Gordon
authors.affiliation: †Department of Chemical and Biological Engineering and ‡Microbiology Doctoral Training Program, University of Wisconsin-Madison, Madison, Wisconsin 53706, United States.
authors.forename: Brian F
authors.initials: BF
authors.lastname: Pfleger
mesh_terms: D001483:Base Sequence; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial / Q000235:genetics; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics; D046940:Synechococcus / Q000235:genetics*; D058615:Synthetic Biology / Q000379:methods; D014805:Vitamin B 12 / Q000235:genetics*
chemical_list: D014805:Vitamin B 12