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

被引:614
作者
Morin, X
Daneman, R
Zavortink, M
Chia, W
机构
[1] Natl Univ Singapore, Inst Mol & Cell Biol, Singapore 117609, Singapore
[2] Univ London Kings Coll, MRC, Ctr Dev Neurobiol, Guys Hosp, London SE1 1UL, England
基金
英国惠康基金;
关键词
D O I
10.1073/pnas.261408198
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
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.
引用
收藏
页码:15050 / 15055
页数:6
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