Synthetic Biology Toolbox for Controlling Gene Expression in the Cyanobacterium Synechococcus sp. strain PCC 7002

Markley, Andrew L.; Begemann, Matthew B.; Clarke, Ryan E.; Gordon, Gina C.; Pfleger, Brian F.

doi:10.1021/sb500260k

Title: Synthetic Biology Toolbox for Controlling Gene Expression in the Cyanobacterium Synechococcus sp. strain PCC 7002

Journal Article · Thu Sep 25 00:00:00 EDT 2014 · ACS Synthetic Biology

DOI:https://doi.org/10.1021/sb500260k· OSTI ID:1158729

Markley, Andrew L. ^[1]; Begemann, Matthew B. ^[1]; Clarke, Ryan E. ^[1]; Gordon, Gina C. ^[1]; Pfleger, Brian F. ^[1]

Department of Chemical and Biological Engineering and ‡Microbiology Doctoral Training Program, University of Wisconsin−Madison, Madison, Wisconsin 53706, United States

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 P_trc constructs. Finally, a RBS library was designed and tested in PCC 7002. The presented synthetic biology toolbox will enable accelerated engineering of PCC 7002.

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Research Organization:: Univ. of Wisconsin, Madison, WI (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER); National Science Foundation (NSF); US Air Force Office of Scientific Research (AFOSR); National Inst. of Health (NIH) (United States)

Grant/Contract Number:: SC0010329; EFRI-1240268; CBET-1149678; SEES 12-15871; FA9550-11-1-0038; NIGMS-5 T32 GM08349

OSTI ID:: 1158729

Alternate ID(s):: OSTI ID: 1454871

Journal Information:: ACS Synthetic Biology, Journal Name: ACS Synthetic Biology Vol. 4 Journal Issue: 5; ISSN 2161-5063

Publisher:: American Chemical SocietyCopyright Statement

Country of Publication:: United States

Language:: English

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Cited by: 128 works

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