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Article: Developing the use of mismatch binding proteins for discovering rare somatic mutations

TitleDeveloping the use of mismatch binding proteins for discovering rare somatic mutations
Authors
Issue Date2005
PublisherAcademic Press. The Journal's web site is located at http://www.elsevier.com/locate/ymcpr
Citation
Molecular And Cellular Probes, 2005, v. 19 n. 3, p. 163-168 How to Cite?
AbstractA method for detecting rare, unknown, somatic mutations could allow presymptomatic cancer screening from human fluids. Immobilized mismatch binding protein can bind DNA heteroduplexes while allowing homoduplexes to be washed away, thus enriching for rare mutations. We examined the potential use, for mutation enrichment, of a fusion protein of maltose binding protein and the mismatch binding protein TaqMutS (MBP-MutS). Unlabeled and fluorescent-labeled oligonucleotides, either perfectly complementary or with single nucleotide mismatches or deletions, were combined to form homo- or heteroduplexes that were then mixed at low ratios of hetero- to homoduplexes and enriched for heteroduplexes. Enrichment was observed using a capillary DNA sequencer. A single base deletion oligonucleotide was enriched by a factor of 29, and a mismatch oligonucleotide was enriched by a factor of 2. N-1 oligonucleotide synthesis fragments were enriched more than were mismatches, suggesting that these deletion fragments may compete for MutS and impede enrichment of mismatches. Purification of oligonucleotides by high pressure liquid chromatography or polyacrylamide gel electrophoresis failed to remove n-1 fragments, thus overcoming this obstacle to enrichment of mismatch mutations may require alternative strategies, such as developing new purification methods or avoiding the use of synthetic oligonucleotides before enrichment. © 2004 Elsevier Ltd. All rights reserved.
Persistent Identifierhttp://hdl.handle.net/10722/162802
ISSN
2015 Impact Factor: 1.565
2015 SCImago Journal Rankings: 0.610
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorBaum, Len_US
dc.contributor.authorNg, Aen_US
dc.contributor.authorLeung, WKen_US
dc.date.accessioned2012-09-05T05:23:43Z-
dc.date.available2012-09-05T05:23:43Z-
dc.date.issued2005en_US
dc.identifier.citationMolecular And Cellular Probes, 2005, v. 19 n. 3, p. 163-168en_US
dc.identifier.issn0890-8508en_US
dc.identifier.urihttp://hdl.handle.net/10722/162802-
dc.description.abstractA method for detecting rare, unknown, somatic mutations could allow presymptomatic cancer screening from human fluids. Immobilized mismatch binding protein can bind DNA heteroduplexes while allowing homoduplexes to be washed away, thus enriching for rare mutations. We examined the potential use, for mutation enrichment, of a fusion protein of maltose binding protein and the mismatch binding protein TaqMutS (MBP-MutS). Unlabeled and fluorescent-labeled oligonucleotides, either perfectly complementary or with single nucleotide mismatches or deletions, were combined to form homo- or heteroduplexes that were then mixed at low ratios of hetero- to homoduplexes and enriched for heteroduplexes. Enrichment was observed using a capillary DNA sequencer. A single base deletion oligonucleotide was enriched by a factor of 29, and a mismatch oligonucleotide was enriched by a factor of 2. N-1 oligonucleotide synthesis fragments were enriched more than were mismatches, suggesting that these deletion fragments may compete for MutS and impede enrichment of mismatches. Purification of oligonucleotides by high pressure liquid chromatography or polyacrylamide gel electrophoresis failed to remove n-1 fragments, thus overcoming this obstacle to enrichment of mismatch mutations may require alternative strategies, such as developing new purification methods or avoiding the use of synthetic oligonucleotides before enrichment. © 2004 Elsevier Ltd. All rights reserved.en_US
dc.languageengen_US
dc.publisherAcademic Press. The Journal's web site is located at http://www.elsevier.com/locate/ymcpren_US
dc.relation.ispartofMolecular and Cellular Probesen_US
dc.subject.meshBase Pair Mismatch - Geneticsen_US
dc.subject.meshChromatography, High Pressure Liquiden_US
dc.subject.meshDna - Genetics - Metabolismen_US
dc.subject.meshDna-Binding Proteins - Metabolismen_US
dc.subject.meshElectrophoresisen_US
dc.subject.meshGene Deletionen_US
dc.subject.meshGenetic Testing - Methodsen_US
dc.subject.meshMutation - Geneticsen_US
dc.titleDeveloping the use of mismatch binding proteins for discovering rare somatic mutationsen_US
dc.typeArticleen_US
dc.identifier.emailLeung, WK:waikleung@hku.hken_US
dc.identifier.authorityLeung, WK=rp01479en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.doi10.1016/j.mcp.2004.11.001en_US
dc.identifier.pmid15797815-
dc.identifier.scopuseid_2-s2.0-15944367458en_US
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-15944367458&selection=ref&src=s&origin=recordpageen_US
dc.identifier.volume19en_US
dc.identifier.issue3en_US
dc.identifier.spage163en_US
dc.identifier.epage168en_US
dc.identifier.isiWOS:000228515500002-
dc.publisher.placeUnited Kingdomen_US
dc.identifier.scopusauthoridBaum, L=7103310839en_US
dc.identifier.scopusauthoridNg, A=55186466600en_US
dc.identifier.scopusauthoridLeung, WK=7201504523en_US

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