Large-scale insertional mutagenesis of a coleopteran stored grain pest, the red flour beetle <it>Tribolium castaneum</it>, identifies embryonic lethal mutations and enhancer traps

<p>Abstract</p> <p>Background</p> <p>Given its sequenced genome and efficient systemic RNA interference response, the red flour beetle <it>Tribolium castaneum </it>is a model organism well suited for reverse genetics. Even so, there is a pressing need for fo...

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Main Authors: Klingler Martin, Beeman Richard W, Shippy Teresa D, Wimmer Ernst A, Lorenzen Marcé D, Schinko Johannes, Trauner Jochen, Bucher Gregor, Brown Susan J
Format: Article
Language:English
Published: BMC 2009-11-01
Series:BMC Biology
Online Access:http://www.biomedcentral.com/1741-7007/7/73
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author Klingler Martin
Beeman Richard W
Shippy Teresa D
Wimmer Ernst A
Lorenzen Marcé D
Schinko Johannes
Trauner Jochen
Bucher Gregor
Brown Susan J
author_facet Klingler Martin
Beeman Richard W
Shippy Teresa D
Wimmer Ernst A
Lorenzen Marcé D
Schinko Johannes
Trauner Jochen
Bucher Gregor
Brown Susan J
author_sort Klingler Martin
collection DOAJ
description <p>Abstract</p> <p>Background</p> <p>Given its sequenced genome and efficient systemic RNA interference response, the red flour beetle <it>Tribolium castaneum </it>is a model organism well suited for reverse genetics. Even so, there is a pressing need for forward genetic analysis to escape the bias inherent in candidate gene approaches.</p> <p>Results</p> <p>To produce easy-to-maintain insertional mutations and to obtain fluorescent marker lines to aid phenotypic analysis, we undertook a large-scale transposon mutagenesis screen. In this screen, we produced more than 6,500 new <it>piggyBac </it>insertions. Of these, 421 proved to be recessive lethal, 75 were semi-lethal, and eight indicated recessive sterility, while 505 showed new enhancer-trap patterns. Insertion junctions were determined for 403 lines and often appeared to be located within transcription units. Insertion sites appeared to be randomly distributed throughout the genome, with the exception of a preference for reinsertion near the donor site.</p> <p>Conclusion</p> <p>A large collection of enhancer-trap and embryonic lethal beetle lines has been made available to the research community and will foster investigations into diverse fields of insect biology, pest control, and evolution. Because the genetic elements used in this screen are species-nonspecific, and because the crossing scheme does not depend on balancer chromosomes, the methods presented herein should be broadly applicable for many insect species.</p>
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spelling doaj.art-a7708765e2c14fa88548262624286f7c2022-12-22T03:00:32ZengBMCBMC Biology1741-70072009-11-01717310.1186/1741-7007-7-73Large-scale insertional mutagenesis of a coleopteran stored grain pest, the red flour beetle <it>Tribolium castaneum</it>, identifies embryonic lethal mutations and enhancer trapsKlingler MartinBeeman Richard WShippy Teresa DWimmer Ernst ALorenzen Marcé DSchinko JohannesTrauner JochenBucher GregorBrown Susan J<p>Abstract</p> <p>Background</p> <p>Given its sequenced genome and efficient systemic RNA interference response, the red flour beetle <it>Tribolium castaneum </it>is a model organism well suited for reverse genetics. Even so, there is a pressing need for forward genetic analysis to escape the bias inherent in candidate gene approaches.</p> <p>Results</p> <p>To produce easy-to-maintain insertional mutations and to obtain fluorescent marker lines to aid phenotypic analysis, we undertook a large-scale transposon mutagenesis screen. In this screen, we produced more than 6,500 new <it>piggyBac </it>insertions. Of these, 421 proved to be recessive lethal, 75 were semi-lethal, and eight indicated recessive sterility, while 505 showed new enhancer-trap patterns. Insertion junctions were determined for 403 lines and often appeared to be located within transcription units. Insertion sites appeared to be randomly distributed throughout the genome, with the exception of a preference for reinsertion near the donor site.</p> <p>Conclusion</p> <p>A large collection of enhancer-trap and embryonic lethal beetle lines has been made available to the research community and will foster investigations into diverse fields of insect biology, pest control, and evolution. Because the genetic elements used in this screen are species-nonspecific, and because the crossing scheme does not depend on balancer chromosomes, the methods presented herein should be broadly applicable for many insect species.</p>http://www.biomedcentral.com/1741-7007/7/73
spellingShingle Klingler Martin
Beeman Richard W
Shippy Teresa D
Wimmer Ernst A
Lorenzen Marcé D
Schinko Johannes
Trauner Jochen
Bucher Gregor
Brown Susan J
Large-scale insertional mutagenesis of a coleopteran stored grain pest, the red flour beetle <it>Tribolium castaneum</it>, identifies embryonic lethal mutations and enhancer traps
BMC Biology
title Large-scale insertional mutagenesis of a coleopteran stored grain pest, the red flour beetle <it>Tribolium castaneum</it>, identifies embryonic lethal mutations and enhancer traps
title_full Large-scale insertional mutagenesis of a coleopteran stored grain pest, the red flour beetle <it>Tribolium castaneum</it>, identifies embryonic lethal mutations and enhancer traps
title_fullStr Large-scale insertional mutagenesis of a coleopteran stored grain pest, the red flour beetle <it>Tribolium castaneum</it>, identifies embryonic lethal mutations and enhancer traps
title_full_unstemmed Large-scale insertional mutagenesis of a coleopteran stored grain pest, the red flour beetle <it>Tribolium castaneum</it>, identifies embryonic lethal mutations and enhancer traps
title_short Large-scale insertional mutagenesis of a coleopteran stored grain pest, the red flour beetle <it>Tribolium castaneum</it>, identifies embryonic lethal mutations and enhancer traps
title_sort large scale insertional mutagenesis of a coleopteran stored grain pest the red flour beetle it tribolium castaneum it identifies embryonic lethal mutations and enhancer traps
url http://www.biomedcentral.com/1741-7007/7/73
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