Engineering of a green‐light inducible gene expression system in Synechocystis sp. PCC6803. Issue 2 (12th December 2013)

• Record Type: Journal Article
• Authors: Abe, Koichi; Miyake, Kotone; Nakamura, Mayumi; Kojima, Katsuhiro; Ferri, Stefano; Ikebukuro, Kazunori; Sode, Koji
• Is Part Of: Microbial biotechnology. Volume 7:Issue 2(2014:Mar.)
• Page Start: 177
• Page End: 183
• Publication Date: 2013-12-12
• Subjects: Microbial biotechnology -- Periodicals; Biotechnology; Microbiology; 660.62
• DOI: 10.1111/1751-7915.12098
• Languages: English
• ISSNs: 1751-7915

Abstract

<abstract abstract-type="main"> <title>Summary</title> <p>In order to construct a green‐light‐regulated gene expression system for cyanobacteria, we characterized a green‐light sensing system derived from <italic>S</italic><italic>ynechocystis</italic> sp. PCC6803, consisting of the green‐light sensing histidine kinase CcaS, the cognate response regulator CcaR, and the promoter of <italic>cpcG2</italic> (P<sub>cpcG</sub><sub>2</sub>). CcaS and CcaR act as a genetic controller and activate gene expression from P<sub>cpcG</sub><sub>2</sub> with green‐light illumination. The green‐light induction level of the native P<sub>cpcG</sub><sub>2</sub> was investigated using GFPuv as a reporter gene inserted in a broad‐host‐range vector. A clear induction of protein expression from native P<sub>cpcG</sub><sub>2</sub> under green‐light illumination was observed; however, the expression level was very low compared with P<sub>trc</sub>, which was reported to act as a constitutive promoter in cyanobacteria. Therefore, a Shine‐Dalgarno‐like sequence derived from the <italic>cpcB</italic> gene was inserted in the 5′ untranslated region of the <italic>cpcG2</italic> gene, and the expression level of CcaR was increased. Thus, constructed engineered green‐light sensing system resulted in about 40‐fold higher protein expression than with the wild‐type promoter with a high ON/OFF ratio under green‐light illumination. The engineered green‐light gene expression system would be a useful genetic tool for controlling gene expression in the emergent cyanobacterial bioprocesses.</p> </abstract>