Plant-adapted green fluorescent protein is a versatile vital reporter for gene expression, protein localization and mitosis in the filamentous fungus, Aspergillus nidulans

被引:167
作者
Fernández-Abalos, JM
Fox, H
Pitt, C
Wells, B
Doonan, JH
机构
[1] John Innes Ctr Plant Sci Res, Norwich NR4 7UH, Norfolk, England
[2] Univ Salamanca, Dept Genet & Microbiol, Salamanca 37007, Spain
基金
英国生物技术与生命科学研究理事会;
关键词
D O I
10.1046/j.1365-2958.1998.00664.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Green fluorescent protein (GFP) is a useful reporter to follow the in vivo behaviour of proteins, but the wildtype gfp gene does not function in many organisms, including many plants and filamentous fungi. We show that codon-modified forms of gfp, produced for use in plants, function effectively in Aspergillus nidulans both as gene expression reporters and as vital reporters for protein location. To demonstrate the use of these modified gfps as reporter genes we have used fluorescence to follow ethanol-induced GFP expression from the alcA promoter. Translational fusions with the modified gfp were used to follow protein location in living cells; plant ER-retention signals targeted GFP to the endoplasmic reticulum, whereas fusion to the GAL4 DNA-binding domain targeted it to the nucleus. Nuclear-targeted GFP allowed real-time observation of nuclear movement and division. These modified gfp genes should provide useful markers to follow gene expression, organelle behaviour and protein trafficking in real time.
引用
收藏
页码:121 / 130
页数:10
相关论文
共 27 条
  • [1] Beckwith S M, 1995, Genet Eng (N Y), V17, P165
  • [2] Virus-mediated delivery of the green fluorescent protein to the endoplasmic reticulum of plant cells
    Boevink, P
    SantaCruz, S
    Hawes, C
    Harris, N
    Oparka, KJ
    [J]. PLANT JOURNAL, 1996, 10 (05) : 935 - 941
  • [3] Characterization of the Aspergillus nidulans aspnd1 gene demonstrates that the ASPND1 antigen, which it encodes, and several Aspergillus fumigatus immunodominant antigens belong to the same family
    Calera, JA
    Ovejero, MC
    LopezMedrano, R
    Segurado, M
    Puente, P
    Leal, F
    [J]. INFECTION AND IMMUNITY, 1997, 65 (04) : 1335 - 1344
  • [4] GREEN FLUORESCENT PROTEIN AS A MARKER FOR GENE-EXPRESSION
    CHALFIE, M
    TU, Y
    EUSKIRCHEN, G
    WARD, WW
    PRASHER, DC
    [J]. SCIENCE, 1994, 263 (5148) : 802 - 805
  • [5] Engineered GFP as a vital reporter in plants
    Chiu, WL
    Niwa, Y
    Zeng, W
    Hirano, T
    Kobayashi, H
    Sheen, J
    [J]. CURRENT BIOLOGY, 1996, 6 (03) : 325 - 330
  • [6] Yeast-enhanced green fluorescent protein (yEGFP): A reporter of gene expression in Candida albicans
    Cormack, BP
    Bertram, G
    Egerton, M
    Gow, NAR
    Falkow, S
    Brown, AJP
    [J]. MICROBIOLOGY-UK, 1997, 143 : 303 - 311
  • [7] UNDERSTANDING, IMPROVING AND USING GREEN FLUORESCENT PROTEINS
    CUBITT, AB
    HEIM, R
    ADAMS, SR
    BOYD, AE
    GROSS, LA
    TSIEN, RY
    [J]. TRENDS IN BIOCHEMICAL SCIENCES, 1995, 20 (11) : 448 - 455
  • [8] DOONAN JH, 1991, J BIOL CHEM, V266, P18889
  • [9] DOONAN JH, 1992, J CELL SCI, V103, P599
  • [10] A NOVEL GENETIC SYSTEM TO DETECT PROTEIN PROTEIN INTERACTIONS
    FIELDS, S
    SONG, OK
    [J]. NATURE, 1989, 340 (6230) : 245 - 246