Engineering of a green-light inducible gene expression system in Synechocystis sp. PCC6803.
Microb Biotechnol
; 7(2): 177-83, 2014 Mar.
Article
em En
| MEDLINE
| ID: mdl-24330635
ABSTRACT
In order to construct a green-light-regulated gene expression system for cyanobacteria, we characterized a green-light sensing system derived from Synechocystis sp. PCC6803, consisting of the green-light sensing histidine kinase CcaS, the cognate response regulator CcaR, and the promoter of cpcG2 (PcpcG 2 ). CcaS and CcaR act as a genetic controller and activate gene expression from PcpcG 2 with green-light illumination. The green-light induction level of the native PcpcG 2 was investigated using GFPuv as a reporter gene inserted in a broad-host-range vector. A clear induction of protein expression from native PcpcG 2 under green-light illumination was observed; however, the expression level was very low compared with Ptrc , which was reported to act as a constitutive promoter in cyanobacteria. Therefore, a Shine-Dalgarno-like sequence derived from the cpcB gene was inserted in the 5' untranslated region of the cpcG2 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.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Regulação Bacteriana da Expressão Gênica
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Ativação Transcricional
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Synechocystis
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Genética Microbiana
Idioma:
En
Revista:
Microb Biotechnol
Ano de publicação:
2014
Tipo de documento:
Article