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Extraction of Venom and Venom Gland Microdissections from Spiders for Proteomic and Transcriptomic Analyses
Venoms are chemically complex secretions typically comprising numerous proteins and peptides with varied physiological activities. Functional characterization of venom proteins has important biomedical applications, including the identification of drug leads or probes for cellular receptors. Spiders...
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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MyJove Corporation
2014
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4353418/ https://www.ncbi.nlm.nih.gov/pubmed/25407635 http://dx.doi.org/10.3791/51618 |
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author | Garb, Jessica E. |
author_facet | Garb, Jessica E. |
author_sort | Garb, Jessica E. |
collection | PubMed |
description | Venoms are chemically complex secretions typically comprising numerous proteins and peptides with varied physiological activities. Functional characterization of venom proteins has important biomedical applications, including the identification of drug leads or probes for cellular receptors. Spiders are the most species rich clade of venomous organisms, but the venoms of only a few species are well-understood, in part due to the difficulty associated with collecting minute quantities of venom from small animals. This paper presents a protocol for the collection of venom from spiders using electrical stimulation, demonstrating the procedure on the Western black widow (Latrodectus hesperus). The collected venom is useful for varied downstream analyses including direct protein identification via mass spectrometry, functional assays, and stimulation of venom gene expression for transcriptomic studies. This technique has the advantage over protocols that isolate venom from whole gland homogenates, which do not separate genuine venom components from cellular proteins that are not secreted as part of the venom. Representative results demonstrate the detection of known venom peptides from the collected sample using mass spectrometry. The venom collection procedure is followed by a protocol for dissecting spider venom glands, with results demonstrating that this leads to the characterization of venom-expressed proteins and peptides at the sequence level. |
format | Online Article Text |
id | pubmed-4353418 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | MyJove Corporation |
record_format | MEDLINE/PubMed |
spelling | pubmed-43534182015-03-12 Extraction of Venom and Venom Gland Microdissections from Spiders for Proteomic and Transcriptomic Analyses Garb, Jessica E. J Vis Exp Genetics Venoms are chemically complex secretions typically comprising numerous proteins and peptides with varied physiological activities. Functional characterization of venom proteins has important biomedical applications, including the identification of drug leads or probes for cellular receptors. Spiders are the most species rich clade of venomous organisms, but the venoms of only a few species are well-understood, in part due to the difficulty associated with collecting minute quantities of venom from small animals. This paper presents a protocol for the collection of venom from spiders using electrical stimulation, demonstrating the procedure on the Western black widow (Latrodectus hesperus). The collected venom is useful for varied downstream analyses including direct protein identification via mass spectrometry, functional assays, and stimulation of venom gene expression for transcriptomic studies. This technique has the advantage over protocols that isolate venom from whole gland homogenates, which do not separate genuine venom components from cellular proteins that are not secreted as part of the venom. Representative results demonstrate the detection of known venom peptides from the collected sample using mass spectrometry. The venom collection procedure is followed by a protocol for dissecting spider venom glands, with results demonstrating that this leads to the characterization of venom-expressed proteins and peptides at the sequence level. MyJove Corporation 2014-11-03 /pmc/articles/PMC4353418/ /pubmed/25407635 http://dx.doi.org/10.3791/51618 Text en Copyright © 2014, Journal of Visualized Experiments http://creativecommons.org/licenses/by-nc-nd/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License. To view a copy of this license, visithttp://creativecommons.org/licenses/by-nc-nd/3.0/ |
spellingShingle | Genetics Garb, Jessica E. Extraction of Venom and Venom Gland Microdissections from Spiders for Proteomic and Transcriptomic Analyses |
title | Extraction of Venom and Venom Gland Microdissections from Spiders for Proteomic and Transcriptomic Analyses |
title_full | Extraction of Venom and Venom Gland Microdissections from Spiders for Proteomic and Transcriptomic Analyses |
title_fullStr | Extraction of Venom and Venom Gland Microdissections from Spiders for Proteomic and Transcriptomic Analyses |
title_full_unstemmed | Extraction of Venom and Venom Gland Microdissections from Spiders for Proteomic and Transcriptomic Analyses |
title_short | Extraction of Venom and Venom Gland Microdissections from Spiders for Proteomic and Transcriptomic Analyses |
title_sort | extraction of venom and venom gland microdissections from spiders for proteomic and transcriptomic analyses |
topic | Genetics |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4353418/ https://www.ncbi.nlm.nih.gov/pubmed/25407635 http://dx.doi.org/10.3791/51618 |
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