Loading…
Persistence of extracellular baculoviral DNA in aquatic microcosms: extraction, purification, and amplification by the polymerase chain reaction (PCR)
Genetically-modified baculoviruses have potential uses as bio-pesticides in forestry. However, the baculoviral occlusion bodies (OBs) may release genetically-modified DNA into the forest environment. In this research, outdoor aquatic microcosms, spiked with 673 μg of genomic DNA (4.4×10 12 target co...
Saved in:
Published in: | Molecular and cellular probes 2005-04, Vol.19 (2), p.75-80 |
---|---|
Main Authors: | , , , , , , |
Format: | Article |
Language: | English |
Subjects: | |
Citations: | Items that this one cites Items that cite this one |
Online Access: | Get full text |
Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
cited_by | cdi_FETCH-LOGICAL-c351t-176ac3fe8c47320ef73fa0496bd127e52c0ad69f4dad62d6657f537b9d5e44103 |
---|---|
cites | cdi_FETCH-LOGICAL-c351t-176ac3fe8c47320ef73fa0496bd127e52c0ad69f4dad62d6657f537b9d5e44103 |
container_end_page | 80 |
container_issue | 2 |
container_start_page | 75 |
container_title | Molecular and cellular probes |
container_volume | 19 |
creator | England, L.S. Pollok, J. Vincent, M. Kreutzweiser, D. Fick, W. Trevors, J.T. Holmes, S.B. |
description | Genetically-modified baculoviruses have potential uses as bio-pesticides in forestry. However, the baculoviral occlusion bodies (OBs) may release genetically-modified DNA into the forest environment. In this research, outdoor aquatic microcosms, spiked with 673
μg of genomic DNA (4.4×10
12 target copies) from the genetically modified baculovirus
Choristoneura fumiferana MNPV
egt
−/
lacZ
+, were exposed to natural summer conditions. A 530
bp DNA fragment from the genome of
CfMNPV
egt
−/
lacZ
+ was detected in field microcosm water samples for about 24
h. The introduced DNA may have persisted for a longer time, but was below the detection limit of the PCR analysis (13.5
pg DNA or 8.9×10
4 target copies ml
−1 water). The detection limit of PCR was determined by spiking water samples with a dilution series of
CfMNPV
egt
−/
lacZ
+ genomic DNA, extracting and purifying the DNA, and then PCR analysis. This study provides some of the first information on the persistence and detection limits of this viral DNA under aquatic ecological conditions, and the methods that can be used to conduct such a molecular-based field study. |
doi_str_mv | 10.1016/j.mcp.2004.09.004 |
format | article |
fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_67395429</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>S0890850804000829</els_id><sourcerecordid>67395429</sourcerecordid><originalsourceid>FETCH-LOGICAL-c351t-176ac3fe8c47320ef73fa0496bd127e52c0ad69f4dad62d6657f537b9d5e44103</originalsourceid><addsrcrecordid>eNp9kc1u1DAUhS1ERaeFB2CDvEIgkXCdOD8uq2oKtFIFFYK15djXqkdJnNpJxbxInxePMoIdqyNb53zSuYeQ1wxyBqz-uMsHPeUFAM9B5EmekQ0DUWeMCf6cbKAVkLUVtKfkLMYdAAgO7Qtyyqq6hQKaDXm6wxBdnHHUSL2l-HsOSmPfL70KtFN66f2jC6qnV98uqRupeljU7DQdnA5e-zjEi2Nodn78QKclOOu0Wl9qNFQNU__3i3Z7Ot8jnXy_HzCoiFTfq8QNuBLou7vtj_cvyYlVfcRXRz0nv758_rm9zm6_f73ZXt5muqzYnLGmVrq02GrelAWgbUqrgIu6M6xosCo0KFMLy02SwtR11diqbDphKuScQXlO3q7cKfiHBeMsBxcP9dWIfomybkpR8UIkI1uNqXWMAa2cghtU2EsG8jCG3Mk0hjyMIUHIJCnz5ghfugHNv8Tx-snwaTVgqvjoMMio3WEJ4wLqWRrv_oP_A5IEnQ4</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>67395429</pqid></control><display><type>article</type><title>Persistence of extracellular baculoviral DNA in aquatic microcosms: extraction, purification, and amplification by the polymerase chain reaction (PCR)</title><source>ScienceDirect Journals</source><creator>England, L.S. ; Pollok, J. ; Vincent, M. ; Kreutzweiser, D. ; Fick, W. ; Trevors, J.T. ; Holmes, S.B.</creator><creatorcontrib>England, L.S. ; Pollok, J. ; Vincent, M. ; Kreutzweiser, D. ; Fick, W. ; Trevors, J.T. ; Holmes, S.B.</creatorcontrib><description>Genetically-modified baculoviruses have potential uses as bio-pesticides in forestry. However, the baculoviral occlusion bodies (OBs) may release genetically-modified DNA into the forest environment. In this research, outdoor aquatic microcosms, spiked with 673
μg of genomic DNA (4.4×10
12 target copies) from the genetically modified baculovirus
Choristoneura fumiferana MNPV
egt
−/
lacZ
+, were exposed to natural summer conditions. A 530
bp DNA fragment from the genome of
CfMNPV
egt
−/
lacZ
+ was detected in field microcosm water samples for about 24
h. The introduced DNA may have persisted for a longer time, but was below the detection limit of the PCR analysis (13.5
pg DNA or 8.9×10
4 target copies ml
−1 water). The detection limit of PCR was determined by spiking water samples with a dilution series of
CfMNPV
egt
−/
lacZ
+ genomic DNA, extracting and purifying the DNA, and then PCR analysis. This study provides some of the first information on the persistence and detection limits of this viral DNA under aquatic ecological conditions, and the methods that can be used to conduct such a molecular-based field study.</description><identifier>ISSN: 0890-8508</identifier><identifier>EISSN: 1096-1194</identifier><identifier>DOI: 10.1016/j.mcp.2004.09.004</identifier><identifier>PMID: 15680207</identifier><language>eng</language><publisher>England: Elsevier Ltd</publisher><subject>Amplification ; Aquatic ; Baculoviridae - genetics ; Detection ; DNA ; DNA, Viral - analysis ; DNA, Viral - genetics ; Ecosystem ; Forest environment ; Forestry ; Fresh Water - chemistry ; Lac Operon ; Persistence</subject><ispartof>Molecular and cellular probes, 2005-04, Vol.19 (2), p.75-80</ispartof><rights>2004 Elsevier Ltd</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c351t-176ac3fe8c47320ef73fa0496bd127e52c0ad69f4dad62d6657f537b9d5e44103</citedby><cites>FETCH-LOGICAL-c351t-176ac3fe8c47320ef73fa0496bd127e52c0ad69f4dad62d6657f537b9d5e44103</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/15680207$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>England, L.S.</creatorcontrib><creatorcontrib>Pollok, J.</creatorcontrib><creatorcontrib>Vincent, M.</creatorcontrib><creatorcontrib>Kreutzweiser, D.</creatorcontrib><creatorcontrib>Fick, W.</creatorcontrib><creatorcontrib>Trevors, J.T.</creatorcontrib><creatorcontrib>Holmes, S.B.</creatorcontrib><title>Persistence of extracellular baculoviral DNA in aquatic microcosms: extraction, purification, and amplification by the polymerase chain reaction (PCR)</title><title>Molecular and cellular probes</title><addtitle>Mol Cell Probes</addtitle><description>Genetically-modified baculoviruses have potential uses as bio-pesticides in forestry. However, the baculoviral occlusion bodies (OBs) may release genetically-modified DNA into the forest environment. In this research, outdoor aquatic microcosms, spiked with 673
μg of genomic DNA (4.4×10
12 target copies) from the genetically modified baculovirus
Choristoneura fumiferana MNPV
egt
−/
lacZ
+, were exposed to natural summer conditions. A 530
bp DNA fragment from the genome of
CfMNPV
egt
−/
lacZ
+ was detected in field microcosm water samples for about 24
h. The introduced DNA may have persisted for a longer time, but was below the detection limit of the PCR analysis (13.5
pg DNA or 8.9×10
4 target copies ml
−1 water). The detection limit of PCR was determined by spiking water samples with a dilution series of
CfMNPV
egt
−/
lacZ
+ genomic DNA, extracting and purifying the DNA, and then PCR analysis. This study provides some of the first information on the persistence and detection limits of this viral DNA under aquatic ecological conditions, and the methods that can be used to conduct such a molecular-based field study.</description><subject>Amplification</subject><subject>Aquatic</subject><subject>Baculoviridae - genetics</subject><subject>Detection</subject><subject>DNA</subject><subject>DNA, Viral - analysis</subject><subject>DNA, Viral - genetics</subject><subject>Ecosystem</subject><subject>Forest environment</subject><subject>Forestry</subject><subject>Fresh Water - chemistry</subject><subject>Lac Operon</subject><subject>Persistence</subject><issn>0890-8508</issn><issn>1096-1194</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2005</creationdate><recordtype>article</recordtype><recordid>eNp9kc1u1DAUhS1ERaeFB2CDvEIgkXCdOD8uq2oKtFIFFYK15djXqkdJnNpJxbxInxePMoIdqyNb53zSuYeQ1wxyBqz-uMsHPeUFAM9B5EmekQ0DUWeMCf6cbKAVkLUVtKfkLMYdAAgO7Qtyyqq6hQKaDXm6wxBdnHHUSL2l-HsOSmPfL70KtFN66f2jC6qnV98uqRupeljU7DQdnA5e-zjEi2Nodn78QKclOOu0Wl9qNFQNU__3i3Z7Ot8jnXy_HzCoiFTfq8QNuBLou7vtj_cvyYlVfcRXRz0nv758_rm9zm6_f73ZXt5muqzYnLGmVrq02GrelAWgbUqrgIu6M6xosCo0KFMLy02SwtR11diqbDphKuScQXlO3q7cKfiHBeMsBxcP9dWIfomybkpR8UIkI1uNqXWMAa2cghtU2EsG8jCG3Mk0hjyMIUHIJCnz5ghfugHNv8Tx-snwaTVgqvjoMMio3WEJ4wLqWRrv_oP_A5IEnQ4</recordid><startdate>20050401</startdate><enddate>20050401</enddate><creator>England, L.S.</creator><creator>Pollok, J.</creator><creator>Vincent, M.</creator><creator>Kreutzweiser, D.</creator><creator>Fick, W.</creator><creator>Trevors, J.T.</creator><creator>Holmes, S.B.</creator><general>Elsevier Ltd</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>20050401</creationdate><title>Persistence of extracellular baculoviral DNA in aquatic microcosms: extraction, purification, and amplification by the polymerase chain reaction (PCR)</title><author>England, L.S. ; Pollok, J. ; Vincent, M. ; Kreutzweiser, D. ; Fick, W. ; Trevors, J.T. ; Holmes, S.B.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c351t-176ac3fe8c47320ef73fa0496bd127e52c0ad69f4dad62d6657f537b9d5e44103</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2005</creationdate><topic>Amplification</topic><topic>Aquatic</topic><topic>Baculoviridae - genetics</topic><topic>Detection</topic><topic>DNA</topic><topic>DNA, Viral - analysis</topic><topic>DNA, Viral - genetics</topic><topic>Ecosystem</topic><topic>Forest environment</topic><topic>Forestry</topic><topic>Fresh Water - chemistry</topic><topic>Lac Operon</topic><topic>Persistence</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>England, L.S.</creatorcontrib><creatorcontrib>Pollok, J.</creatorcontrib><creatorcontrib>Vincent, M.</creatorcontrib><creatorcontrib>Kreutzweiser, D.</creatorcontrib><creatorcontrib>Fick, W.</creatorcontrib><creatorcontrib>Trevors, J.T.</creatorcontrib><creatorcontrib>Holmes, S.B.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Molecular and cellular probes</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>England, L.S.</au><au>Pollok, J.</au><au>Vincent, M.</au><au>Kreutzweiser, D.</au><au>Fick, W.</au><au>Trevors, J.T.</au><au>Holmes, S.B.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Persistence of extracellular baculoviral DNA in aquatic microcosms: extraction, purification, and amplification by the polymerase chain reaction (PCR)</atitle><jtitle>Molecular and cellular probes</jtitle><addtitle>Mol Cell Probes</addtitle><date>2005-04-01</date><risdate>2005</risdate><volume>19</volume><issue>2</issue><spage>75</spage><epage>80</epage><pages>75-80</pages><issn>0890-8508</issn><eissn>1096-1194</eissn><abstract>Genetically-modified baculoviruses have potential uses as bio-pesticides in forestry. However, the baculoviral occlusion bodies (OBs) may release genetically-modified DNA into the forest environment. In this research, outdoor aquatic microcosms, spiked with 673
μg of genomic DNA (4.4×10
12 target copies) from the genetically modified baculovirus
Choristoneura fumiferana MNPV
egt
−/
lacZ
+, were exposed to natural summer conditions. A 530
bp DNA fragment from the genome of
CfMNPV
egt
−/
lacZ
+ was detected in field microcosm water samples for about 24
h. The introduced DNA may have persisted for a longer time, but was below the detection limit of the PCR analysis (13.5
pg DNA or 8.9×10
4 target copies ml
−1 water). The detection limit of PCR was determined by spiking water samples with a dilution series of
CfMNPV
egt
−/
lacZ
+ genomic DNA, extracting and purifying the DNA, and then PCR analysis. This study provides some of the first information on the persistence and detection limits of this viral DNA under aquatic ecological conditions, and the methods that can be used to conduct such a molecular-based field study.</abstract><cop>England</cop><pub>Elsevier Ltd</pub><pmid>15680207</pmid><doi>10.1016/j.mcp.2004.09.004</doi><tpages>6</tpages></addata></record> |
fulltext | fulltext |
identifier | ISSN: 0890-8508 |
ispartof | Molecular and cellular probes, 2005-04, Vol.19 (2), p.75-80 |
issn | 0890-8508 1096-1194 |
language | eng |
recordid | cdi_proquest_miscellaneous_67395429 |
source | ScienceDirect Journals |
subjects | Amplification Aquatic Baculoviridae - genetics Detection DNA DNA, Viral - analysis DNA, Viral - genetics Ecosystem Forest environment Forestry Fresh Water - chemistry Lac Operon Persistence |
title | Persistence of extracellular baculoviral DNA in aquatic microcosms: extraction, purification, and amplification by the polymerase chain reaction (PCR) |
url | http://sfxeu10.hosted.exlibrisgroup.com/loughborough?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-01T00%3A12%3A30IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Persistence%20of%20extracellular%20baculoviral%20DNA%20in%20aquatic%20microcosms:%20extraction,%20purification,%20and%20amplification%20by%20the%20polymerase%20chain%20reaction%20(PCR)&rft.jtitle=Molecular%20and%20cellular%20probes&rft.au=England,%20L.S.&rft.date=2005-04-01&rft.volume=19&rft.issue=2&rft.spage=75&rft.epage=80&rft.pages=75-80&rft.issn=0890-8508&rft.eissn=1096-1194&rft_id=info:doi/10.1016/j.mcp.2004.09.004&rft_dat=%3Cproquest_cross%3E67395429%3C/proquest_cross%3E%3Cgrp_id%3Ecdi_FETCH-LOGICAL-c351t-176ac3fe8c47320ef73fa0496bd127e52c0ad69f4dad62d6657f537b9d5e44103%3C/grp_id%3E%3Coa%3E%3C/oa%3E%3Curl%3E%3C/url%3E&rft_id=info:oai/&rft_pqid=67395429&rft_id=info:pmid/15680207&rfr_iscdi=true |