title: A synthetic gene coding for the green fluorescent protein (GFP) is a versatile reporter in Chlamydomonas reinhardtii.
abstract: The use of Chlamydomonas reinhardtii as a model system has been hindered by difficulties encountered in expressing foreign genes. We have synthesised a gene encoding green fluorescent protein (GFP) adapted to the codon usage of C. reinhardtii (cgfp). After verifying the gene was functional in Escherichia coli, the cgfp was fused in frame to the phleomycin resistance gene ble from Streptoalloteichus hindustanus and expressed in C. reinhardtii under control of the rbcS2 promoter and intron sequences. The GFP-fluorescence was seen only in the nucleus demonstrating the nuclear accumulation of the Ble-GFP fusion protein. The cgfp was also fused to the chlamyopsin gene, cop, and expressed in C. reinhardtii under control of the cop promoter. The eyespot became fluorescent indicating that the opsin-GFP fusion protein was correctly directed into the eyespot along with the endogenous unmodified opsin. We conclude that cgfp provides a useful tool to visualize protein synthesis and localisation in vivo in C. reinhardtii and possibly in related green algal species.
authors.affiliation: Institut für Biochemie, Genetik und Mikrobiologie, Universität Regensburg, Germany.
authors.forename: M
authors.initials: M
authors.lastname: Fuhrmann
authors.forename: W
authors.initials: W
authors.lastname: Oertel
authors.forename: P
authors.initials: P
authors.lastname: Hegemann
mesh_terms: D020418:Algal Proteins; D000595:Amino Acid Sequence; D000818:Animals; D001059:Apoproteins / Q000096:biosynthesis / Q000235:genetics; D001483:Base Sequence; D002352:Carrier Proteins / Q000096:biosynthesis / Q000235:genetics; D016825:Chlamydomonas reinhardtii / Q000235:genetics* / Q000378:metabolism; D004274:DNA, Recombinant / Q000235:genetics; D015870:Gene Expression; D017930:Genes, Reporter; D005813:Genes, Synthetic; D049452:Green Fluorescent Proteins; D047908:Intracellular Signaling Peptides and Proteins; D008164:Luminescent Proteins / Q000096:biosynthesis / Q000235:genetics*; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D014170:Transformation, Genetic
chemical_list: D020418:Algal Proteins; D001059:Apoproteins; D002352:Carrier Proteins; D004274:DNA, Recombinant; D047908:Intracellular Signaling Peptides and Proteins; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; C097338:chlamyopsin protein, Chlamydomonas reinhardtii; D049452:Green Fluorescent Proteins