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Proc Natl Acad Sci U S A. 2002 Jul 23;99(15):9948-53. Epub 2002 Jul 2.

A deletion-generator compound element allows deletion saturation analysis for genomewide phenotypic annotation.

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1
Department of Molecular and Cellular Biology, Harvard University, 16 Divinity Avenue, Cambridge, MA 02138, USA.

Abstract

With the available eukaryotic genome sequences, there are predictions of thousands of previously uncharacterized genes without known function or available mutational variant. Thus, there is an urgent need for efficient genetic tools for genomewide phenotypic analysis. Here we describe such a tool: a deletion-generator technology that exploits properties of a double transposable element to produce molecularly defined deletions at high density and with high efficiency. This double element, called P[wHy], is composed of a "deleter" element hobo, bracketed by two genetic markers and inserted into a "carrier" P element. We have used this P[wHy] element in Drosophila melanogaster to generate sets of nested deletions of sufficient coverage to discriminate among every transcription unit within 60 kb of the starting insertion site. Because these two types of mobile elements, carrier and deleter, can be found in other species, our strategy should be applicable to phenotypic analysis in a variety of model organisms.

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PMID:
12096187
PMCID:
PMC126605
DOI:
10.1073/pnas.142310099
[Indexed for MEDLINE]
Free PMC Article

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