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Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, CA 94305-5402, USA
Molecular and Cell Biology, University of Aberdeen, Institute of Medical Sciences, Foresterhill, Aberdeen AB25 2ZD, UK
Zeneca Pharmaceuticals, Alderley Park, Macclesfield SK10 4TG, UK
Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, Hamilton, MT 59840, USA
6(European requests for plasmid to Alistair J. P. Brown, e-mail: al.brownt@abdn.ac.uk; all other requests to Brendan P. Cormack)
5Author for correspondence: Brendan P. Cormack. e-mail: cormack@cmgm.stanford.edu
ABSTRACT
The green fluorescent protein (GFP) of Aequorea victoria has been developed here as a reporter for gene expression and protein localization in Candida albicans. When wild-type (wt) GFP was expressed in C. albicans, it was not possible to detect fluorescence or a translation product for the wt protein. Since this was probably due in part to the presence of the non-canonical CTG serine codon in the Aequorea sequence, this codon was changed to the leucine codon TTG. C. albicans cells expressing this construct contained GFP mRNA but were non-fluorescent and contained no detectable translation product. Hence a codon-optimized GFP gene was constructed in which all of the 239 amino acids are encoded by optimal codons for C albicans. In this gene were also incorporated two previously identified mutations in the chromophore that increase GFP fluorescence. C. albicans cells expressing this yeast-enhanced GFP gene (yEGFP3) are fluorescent and contain GFP protein. yEGFP3 can be used as a versatile reporter of gene expression in C. albicans and Saccharomyces cerevisiae and the optimized GFP described here should have broad applications in these and other fungal species.
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E. B. Vervoort, A. v. Ravestein, N. N. M. E. v. Peij, J. C. Heikoop, P. J. M. v. Haastert, G. F. Verheijden, and M. H. K. Linskens Optimizing heterologous expression in Dictyostelium: importance of 5' codon adaptation Nucleic Acids Res., May 15, 2000; 28(10): 2069 - 2074. [Abstract] [Full Text] [PDF] |
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N. Ramanan and Y. Wang A High-Affinity Iron Permease Essential for Candida albicans Virulence Science, May 12, 2000; 288(5468): 1062 - 1064. [Abstract] [Full Text] |
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J. Chen and P. Ahlquist Brome Mosaic Virus Polymerase-Like Protein 2a Is Directed to the Endoplasmic Reticulum by Helicase-Like Viral Protein 1a J. Virol., May 1, 2000; 74(9): 4310 - 4318. [Abstract] [Full Text] |
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T.-W. Soong, T.-F. Yong, N. Ramanan, and Y. Wang The Candida albicans antiporter gene CNH1 has a role in Na+ and H+ transport, salt tolerance, and morphogenesis Microbiology, May 1, 2000; 146(5): 1035 - 1044. [Abstract] [Full Text] |
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C. R. DeMattei, C. P. Davis, and J. B. Konopka Point Mutations Identify a Conserved Region of the Saccharomyces cerevisiae AFR1 Gene That Is Essential for Both the Pheromone Signaling and Morphogenesis Functions Genetics, May 1, 2000; 155(1): 43 - 55. [Abstract] [Full Text] |
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A. Alvarez-Barrientos, J. Arroyo, R. Canton, C. Nombela, and M. Sanchez-Perez Applications of Flow Cytometry to Clinical Microbiology Clin. Microbiol. Rev., April 1, 2000; 13(2): 167 - 195. [Abstract] [Full Text] [PDF] |
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Z. Weissman, I. Berdicevsky, B.-Z. Cavari, and D. Kornitzer The high copper tolerance of Candida albicans is mediated by a P-type ATPase PNAS, March 28, 2000; 97(7): 3520 - 3525. [Abstract] [Full Text] [PDF] |
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C. L. Gordon, V. Khalaj, A. F. J. Ram, D. B. Archer, J. L. Brookman, A. P. J. Trinci, D. J. Jeenes, J. H. Doonan, B. Wells, P. J. Punt, et al. Glucoamylase::green fluorescent protein fusions to monitor protein secretion in Aspergillus niger Microbiology, February 1, 2000; 146(2): 415 - 426. [Abstract] [Full Text] |
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S. V. Avery, S. Malkapuram, C. Mateus, and K. S. Babb Copper/Zinc-Superoxide Dismutase Is Required for Oxytetracycline Resistance of Saccharomyces cerevisiae J. Bacteriol., January 1, 2000; 182(1): 76 - 80. [Abstract] [Full Text] |
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L. Rubino, A. Di Franco, and M. Russo Expression of a plant virus non-structural protein in Saccharomyces cerevisiae causes membrane proliferation and altered mitochondrial morphology J. Gen. Virol., January 1, 2000; 81(1): 279 - 286. [Abstract] [Full Text] |
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J. L. Urbanowski and R. C. Piper The Iron Transporter Fth1p Forms a Complex with the Fet5 Iron Oxidase and Resides on the Vacuolar Membrane J. Biol. Chem., December 31, 1999; 274(53): 38061 - 38070. [Abstract] [Full Text] [PDF] |
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A. Madania, P. Dumoulin, S. Grava, H. Kitamoto, C. Schärer-Brodbeck, A. Soulard, V. Moreau, and B. Winsor The Saccharomyces cerevisiae Homologue of Human Wiskott-Aldrich Syndrome Protein Las17p Interacts with the Arp2/3 Complex Mol. Biol. Cell, October 1, 1999; 10(10): 3521 - 3538. [Abstract] [Full Text] |
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Y. C. Chang and K. J. Kwon-Chung Isolation, Characterization, and Localization of a Capsule-Associated Gene, CAP10, of Cryptococcus neoformans J. Bacteriol., September 15, 1999; 181(18): 5636 - 5643. [Abstract] [Full Text] |
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A. B. Herrero, M. C. Lopez, S. Garcia, A. Schmidt, F. Spaltmann, J. Ruiz-Herrera, and A. Dominguez Control of Filament Formation in Candida albicans by Polyamine Levels Infect. Immun., September 1, 1999; 67(9): 4870 - 4878. [Abstract] [Full Text] [PDF] |
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C. M. Hull and A. D. Johnson Identification of a Mating Type-Like Locus in the Asexual Pathogenic Yeast Candida albicans Science, August 20, 1999; 285(5431): 1271 - 1275. [Abstract] [Full Text] |
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J. S. Tirnauer, E. O'Toole, L. Berrueta, B. E. Bierer, and D. Pellman Yeast Bim1p Promotes the G1-specific Dynamics of Microtubules J. Cell Biol., May 31, 1999; 145(5): 993 - 1007. [Abstract] [Full Text] [PDF] |
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M. del Poeta, D. L. Toffaletti, T. H. Rude, S. D. Sparks, J. Heitman, and J. R. Perfect Cryptococcus neoformans Differential Gene Expression Detected In Vitro and In Vivo with Green Fluorescent Protein Infect. Immun., April 1, 1999; 67(4): 1812 - 1820. [Abstract] [Full Text] [PDF] |
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A. Nern and R. A. Arkowitz A Cdc24p-Far1p-G{beta}{gamma} Protein Complex Required for Yeast Orientation during Mating J. Cell Biol., March 22, 1999; 144(6): 1187 - 1202. [Abstract] [Full Text] [PDF] |
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H. K. Edskes, V. T. Gray, and R. B. Wickner The [URE3] prion is an aggregated form of Ure2p that can be cured by overexpression of Ure2p fragments PNAS, February 16, 1999; 96(4): 1498 - 1503. [Abstract] [Full Text] [PDF] |
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A Balguerie, P Sivadon, M Bonneu, and M Aigle Rvs167p, the budding yeast homolog of amphiphysin, colocalizes with actin patches J. Cell Sci., January 8, 1999; 112(15): 2529 - 2537. [Abstract] [PDF] |
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H. J. Chial, M. P. Rout, T. H. Giddings Jr., and M. Winey Saccharomyces cerevisiae Ndc1p Is a Shared Component of Nuclear Pore Complexes and Spindle Pole Bodies J. Cell Biol., December 28, 1998; 143(7): 1789 - 1800. [Abstract] [Full Text] [PDF] |
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