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Nucleic Acids Res. 2010 May;38(8):2577-93. doi: 10.1093/nar/gkq164. Epub 2010 Mar 17.

Design and characterization of molecular tools for a Synthetic Biology approach towards developing cyanobacterial biotechnology.

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  • 1Department of Photochemistry and Molecular Science, Angström Laboratories, Uppsala University, P.O. Box 523, SE-751 20 Uppsala, Sweden.

Abstract

Cyanobacteria are suitable for sustainable, solar-powered biotechnological applications. Synthetic biology connects biology with computational design and an engineering perspective, but requires efficient tools and information about the function of biological parts and systems. To enable the development of cyanobacterial Synthetic Biology, several molecular tools were developed and characterized: (i) a broad-host-range BioBrick shuttle vector, pPMQAK1, was constructed and confirmed to replicate in Escherichia coli and three different cyanobacterial strains. (ii) The fluorescent proteins Cerulean, GFPmut3B and EYFP have been demonstrated to work as reporter proteins in cyanobacteria, in spite of the strong background of photosynthetic pigments. (iii) Several promoters, like P(rnpB) and variants of P(rbcL), and a version of the promoter P(trc) with two operators for enhanced repression, were developed and characterized in Synechocystis sp. strain PCC6803. (iv) It was shown that a system for targeted protein degradation, which is needed to enable dynamic expression studies, is working in Synechocystis sp. strain PCC6803. The pPMQAK1 shuttle vector allows the use of the growing numbers of BioBrick parts in many prokaryotes, and the other tools herein implemented facilitate the development of new parts and systems in cyanobacteria.

PMID:
20236988
PMCID:
PMC2860132
DOI:
10.1093/nar/gkq164
[PubMed - indexed for MEDLINE]
Free PMC Article
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