title: Engineering of a green-light inducible gene expression system in Synechocystis sp. PCC6803.
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.
authors: Department of Biotechnology and Life Science, Tokyo University of Agriculture & Technology, 2-24-16 Naka-cho, Koganei, Tokyo, 184-8588, Japan; JST, CREST, 2-24-16 Naka-cho, Koganei, Tokyo, 184-8588, Japan.
authors: Koichi
authors: K
authors: Abe
authors: Kotone
authors: K
authors: Miyake
authors: Mayumi
authors: M
authors: Nakamura
authors: Katsuhiro
authors: K
authors: Kojima
authors: Stefano
authors: S
authors: Ferri
authors: Kazunori
authors: K
authors: Ikebukuro
authors: Koji
authors: K
authors: Sode
mesh_terms: D019677:Artificial Gene Fusion; D001426:Bacterial Proteins / Q000235:genetics; D015870:Gene Expression; D015964:Gene Expression Regulation, Bacterial / Q000528:radiation effects*; D017930:Genes, Reporter; D005818:Genetic Engineering; D005827:Genetics, Microbial / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D008027:Light; D011401:Promoter Regions, Genetic; D046939:Synechocystis / Q000235:genetics* / Q000528:radiation effects*; D015533:Transcriptional Activation / Q000528:radiation effects*
chemical_list: D001426:Bacterial Proteins; D049452:Green Fluorescent Proteins