Literature DB >> 11742088

A protein trap strategy to detect GFP-tagged proteins expressed from their endogenous loci in Drosophila.

X Morin1, R Daneman, M Zavortink, W Chia.   

Abstract

In Drosophila, enhancer trap strategies allow rapid access to expression patterns, molecular data, and mutations in trapped genes. However, they do not give any information at the protein level, e.g., about the protein subcellular localization. Using the green fluorescent protein (GFP) as a mobile artificial exon carried by a transposable P-element, we have developed a protein trap system. We screened for individual flies, in which GFP tags full-length endogenous proteins expressed from their endogenous locus, allowing us to observe their cellular and subcellular distribution. GFP fusions are targeted to virtually any compartment of the cell. In the case of insertions in previously known genes, we observe that the subcellular localization of the fusion protein corresponds to the described distribution of the endogenous protein. The artificial GFP exon does not disturb upstream and downstream splicing events. Many insertions correspond to genes not predicted by the Drosophila Genome Project. Our results show the feasibility of a protein trap in Drosophila. GFP reveals in real time the dynamics of protein's distribution in the whole, live organism and provides useful markers for a number of cellular structures and compartments.

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Year:  2001        PMID: 11742088      PMCID: PMC64981          DOI: 10.1073/pnas.261408198

Source DB:  PubMed          Journal:  Proc Natl Acad Sci U S A        ISSN: 0027-8424            Impact factor:   11.205


  32 in total

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Journal:  Genes Dev       Date:  2000-06-01       Impact factor: 11.361

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Authors:  S M Mount; C Burks; G Hertz; G D Stormo; O White; C Fields
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Authors:  W C Skarnes; B A Auerbach; A L Joyner
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4.  Searching for pattern and mutation in the Drosophila genome with a P-lacZ vector.

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Journal:  Genes Dev       Date:  1989-09       Impact factor: 11.361

5.  P-element-mediated enhancer detection: an efficient method for isolating and characterizing developmentally regulated genes in Drosophila.

Authors:  C Wilson; R K Pearson; H J Bellen; C J O'Kane; U Grossniklaus; W J Gehring
Journal:  Genes Dev       Date:  1989-09       Impact factor: 11.361

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8.  Suboptimal 5' and 3' splice sites regulate alternative splicing of Drosophila melanogaster myosin heavy chain transcripts in vitro.

Authors:  D Hodges; S I Bernstein
Journal:  Mech Dev       Date:  1992-05       Impact factor: 1.882

9.  A transgene containing lacZ is expressed in primary sensory neurons in zebrafish.

Authors:  T A Bayer; J A Campos-Ortega
Journal:  Development       Date:  1992-06       Impact factor: 6.868

10.  Targeted gene expression as a means of altering cell fates and generating dominant phenotypes.

Authors:  A H Brand; N Perrimon
Journal:  Development       Date:  1993-06       Impact factor: 6.868

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  391 in total

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Journal:  J Genet       Date:  2010-12       Impact factor: 1.166

4.  Green fluorescent protein tagging Drosophila proteins at their native genomic loci with small P elements.

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Journal:  Genetics       Date:  2003-11       Impact factor: 4.562

5.  piggyBac-based insertional mutagenesis in the presence of stably integrated P elements in Drosophila.

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Journal:  Proc Natl Acad Sci U S A       Date:  2003-06-11       Impact factor: 11.205

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7.  Quantitative GFP fluorescence as an indicator of recombinant protein synthesis in transgenic plants.

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Journal:  Plant Cell Rep       Date:  2003-07-04       Impact factor: 4.570

8.  Stringent analysis of gene function and protein-protein interactions using fluorescently tagged genes.

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Journal:  Genetics       Date:  2011-12-14       Impact factor: 4.562

9.  Non-autonomous modulation of heart rhythm, contractility and morphology in adult fruit flies.

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10.  Flytrap, a database documenting a GFP protein-trap insertion screen in Drosophila melanogaster.

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Journal:  Nucleic Acids Res       Date:  2004-01-01       Impact factor: 16.971

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