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Peptides
Volume 28, Issue 1, January 2007, Pages 51-56
Invertebrate Neuropeptides VII, Invertebrate Neuropeptide Conference 2006
 
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doi:10.1016/j.peptides.2006.08.026    
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Copyright © 2006 Elsevier Inc. All rights reserved.

A model genetic system for testing the in vivo function of peptide toxins

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Hugo W. Tedforda, 1, Francesco Maggioa, 2, Robert A. Reenanb, Corresponding Author Contact Information, E-mail The Corresponding Author and Glenn Kinga, Corresponding Author Contact Information, E-mail The Corresponding Author

aDepartment of Molecular, Microbial & Structural Biology, University of Connecticut Health Center, 263 Farmington Avenue, Farmington, CT 06030, USA

bDepartment of Genetics & Developmental Biology, University of Connecticut Health Center, 263 Farmington Avenue, Farmington, CT 06030, USA


Received 6 April 2006; 
revised 8 August 2006; 
accepted 8 August 2006. 
Available online 1 December 2006.

Abstract

We have developed a model genetic system for analyzing the function of peptide toxins from animal venoms. We engineered and propagated strains of Drosophila melanogaster expressing heat-inducible transgenes encoding either κ-ACTX-Hv1c or ω-ACTX-Hv1a, two insect-specific neurotoxic peptides found in the venom of the Australian funnel-web spider Hadronyche versuta. Heat induction of transgene expression for 20 min was sufficient to kill all transgenic flies, indicating that the ion channels targeted by these toxins are viable insecticide targets. The unusual phenotype of flies induced to express ω-ACTX-Hv1a recapitulates that of a hypomorphic allele of the high-voltage-activated calcium channel Dmca1D, suggesting that this is likely to be the target of ω-ACTX-Hv1a.

Keywords: Peptide toxin; Atracotoxin; Drosophila melanogaster; Transgenesis

Article Outline

1. Introduction
2. Experimental
2.1. Choice of signal sequence
2.2. Construction of transformation vectors encoding ω-ACTX-Hv1a and κ-ACTX-Hv1c
2.3. Propagation of transgenic fly lines
2.4. Timecourse of κ-ACTX-Hv1c induction
3. Results
3.1. Response of flies to induction of κ-ACTX-Hv1c transgene expression
3.2. Response of flies to induction of ω-ACTX-Hv1a transgene expression
3.3. Mating of flies expressing a κ-ACTX-Hv1c transgene
4. Discussion
4.1. An in vivo model system for studying peptidic neurotoxins
4.2. Potential modifications and uses of the inducible transgenic toxin system
4.3. Use of peptide toxins in biological insect control
Acknowledgements
References



Corresponding Author Contact InformationCorresponding author. Tel.: +1 860 679 3691.
Corresponding Author Contact InformationCorresponding author at: Departments of Molecular, Microbial & Structural Biology, University of Connecticut Health Center, 263 Farmington Avenue, Farmington, CT 06030, USA. Tel.: +1 860 679 8364; fax: +1 860 679 1652.
1 Present address: Hotchkiss Brain Institute, Department of Physiology and Biophysics, University of Calgary, Calgary, Alberta T2N 4N1, Canada.
2 Present address: Bristol-Myers Squibb, 6000 Thompson Road, East Syracuse, NY 13057, USA.

Peptides
Volume 28, Issue 1, January 2007, Pages 51-56
Invertebrate Neuropeptides VII, Invertebrate Neuropeptide Conference 2006
 
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