title: Modular generation of fluorescent phycobiliproteins.
abstract: Phycobiliproteins are brightly-fluorescent light-harvesting pigments for photosynthesis in cyanobacteria and red algae. They are also of interest as fluorescent biomarkers, but their heterologous generation in vivo has previously required multiple transformations. We report here a modular approach that requires only two DNA segments. The first codes for the apo-protein. The second codes for fusions capable of chromophore biosynthesis and its covalent attachment to the apo-protein; it contains the genes of heme oxygenase, a bilin reductase, and a chromophore lyase. Phycobiliproteins containing phycoerythrobilin (λ(fluor) ~ 560 nm), phycourobilin (λ(fluor) ~ 500 nm), phycocyanobilin (λ(fluor) ~ 630 nm) or phycoviolobilin (λ(fluor) ~ 580 nm) were obtained in high yield in E. coli. This approach facilitates chromophorylation studies of phycobiliproteins, as well as their use for fluorescence labeling based on their high fluorescence.
authors.affiliation: College of Environmental Science and Engineering, Huazhong University of Science and Technology, Wuhan 430074, P R China.
authors.forename: Xian-Jun
authors.initials: XJ
authors.lastname: Wu
authors.forename: Kun
authors.initials: K
authors.lastname: Chang
authors.forename: Juan
authors.initials: J
authors.lastname: Luo
authors.forename: Ming
authors.initials: M
authors.lastname: Zhou
authors.forename: Hugo
authors.initials: H
authors.lastname: Scheer
authors.forename: Kai-Hong
authors.initials: KH
authors.lastname: Zhao
mesh_terms: D003001:Cloning, Molecular; D000458:Cyanobacteria / Q000737:chemistry / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D005456:Fluorescent Dyes / Q000737:chemistry* / Q000378:metabolism; D052979:Phycobiliproteins / Q000737:chemistry* / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics; D000461:Rhodophyta / Q000737:chemistry / Q000235:genetics*
chemical_list: D005456:Fluorescent Dyes; D052979:Phycobiliproteins; D011993:Recombinant Fusion Proteins