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Article: Identification of factors influencing strand bias in oligonucleotide-mediated recombination in Escherichia coli

TitleIdentification of factors influencing strand bias in oligonucleotide-mediated recombination in Escherichia coli
Authors
Issue Date2003
PublisherOxford University Press. The Journal's web site is located at http://nar.oxfordjournals.org/
Citation
Nucleic Acids Research, 2003, v. 31 n. 22, p. 6674-6687 How to Cite?
AbstractRecombinogenic engineering methodology, also known as recombineering, utilizes homologous recombination to create targeted changes in cellular DNA with great specificity and flexibility. In Escherichia coli, the Red recombination system from bacteriophage lambda has been used successfully to modify both plasmid and chromosomal DNA in a highly efficient manner, using either a linear double-stranded DNA fragment or a synthetic single-stranded oligonucleotide (SSO). The current model for Red/SSO-mediated recombination involves the SSO first annealing to a transient, single-stranded region of DNA before being incorporated into the chromosome or plasmid target. It has been observed previously, in both eukaryotes and prokaryotes, that mutations in the two strands of the DNA double helix are 'corrected' by complementary SSOs with differing efficiencies. Here we investigate further the factors that influence the strand bias as well as the overall efficiency of Red/ SSO-mediated recombination in E.coli. We show that the direction of DNA replication and the nature of the SSO-encoded mismatch are the main factors dictating the recombinational strand bias. However, the influence that the SSO-encoded mismatch exerts upon the recombinational strand bias is abolished in E.coli strains that are defective in mismatch repair (MMR). This reflects the fact that different base-base mispairs are corrected by the mutS/H/L-dependent MMR pathway with differing efficiencies. Furthermore, our data indicate that transcription has negligible influence on the strand bias. These results demonstrate for the first time that the interplay between DNA replication and MMR has a major effect on the efficiency and strand bias of Red/SSO-mediated recombination in E.coli.
Persistent Identifierhttp://hdl.handle.net/10722/48987
ISSN
2015 Impact Factor: 9.202
2015 SCImago Journal Rankings: 7.458
PubMed Central ID
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorLi, XTen_HK
dc.contributor.authorCostantino, Nen_HK
dc.contributor.authorLu, LYen_HK
dc.contributor.authorLiu, DPen_HK
dc.contributor.authorWatt, RMen_HK
dc.contributor.authorCheah, KSEen_HK
dc.contributor.authorCourt, DLen_HK
dc.contributor.authorHuang, JDen_HK
dc.date.accessioned2008-06-12T06:31:26Z-
dc.date.available2008-06-12T06:31:26Z-
dc.date.issued2003en_HK
dc.identifier.citationNucleic Acids Research, 2003, v. 31 n. 22, p. 6674-6687en_HK
dc.identifier.issn0305-1048en_HK
dc.identifier.urihttp://hdl.handle.net/10722/48987-
dc.description.abstractRecombinogenic engineering methodology, also known as recombineering, utilizes homologous recombination to create targeted changes in cellular DNA with great specificity and flexibility. In Escherichia coli, the Red recombination system from bacteriophage lambda has been used successfully to modify both plasmid and chromosomal DNA in a highly efficient manner, using either a linear double-stranded DNA fragment or a synthetic single-stranded oligonucleotide (SSO). The current model for Red/SSO-mediated recombination involves the SSO first annealing to a transient, single-stranded region of DNA before being incorporated into the chromosome or plasmid target. It has been observed previously, in both eukaryotes and prokaryotes, that mutations in the two strands of the DNA double helix are 'corrected' by complementary SSOs with differing efficiencies. Here we investigate further the factors that influence the strand bias as well as the overall efficiency of Red/ SSO-mediated recombination in E.coli. We show that the direction of DNA replication and the nature of the SSO-encoded mismatch are the main factors dictating the recombinational strand bias. However, the influence that the SSO-encoded mismatch exerts upon the recombinational strand bias is abolished in E.coli strains that are defective in mismatch repair (MMR). This reflects the fact that different base-base mispairs are corrected by the mutS/H/L-dependent MMR pathway with differing efficiencies. Furthermore, our data indicate that transcription has negligible influence on the strand bias. These results demonstrate for the first time that the interplay between DNA replication and MMR has a major effect on the efficiency and strand bias of Red/SSO-mediated recombination in E.coli.en_HK
dc.format.extent386 bytes-
dc.format.mimetypetext/html-
dc.languageengen_HK
dc.publisherOxford University Press. The Journal's web site is located at http://nar.oxfordjournals.org/en_HK
dc.relation.ispartofNucleic Acids Researchen_HK
dc.rightsCreative Commons: Attribution 3.0 Hong Kong License-
dc.subject.meshDNA Repair Enzymesen_HK
dc.subject.meshEscherichia coli - geneticsen_HK
dc.subject.meshOligonucleotides - genetics - metabolismen_HK
dc.subject.meshRecombination, Geneticen_HK
dc.subject.meshSignal Transduction - geneticsen_HK
dc.titleIdentification of factors influencing strand bias in oligonucleotide-mediated recombination in Escherichia colien_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=0305-1048&volume=31&issue=22&spage=6674&epage=6687&date=2003&atitle=Identification+of+factors+influencing+strand+bias+in+oligonucleotide-mediated+recombination+in+Escherichia+colien_HK
dc.identifier.emailWatt, RM:rmwatt@hku.hken_HK
dc.identifier.emailCheah, KSE:hrmbdkc@hku.hken_HK
dc.identifier.emailHuang, JD:jdhuang@hkucc.hku.hken_HK
dc.identifier.authorityWatt, RM=rp00043en_HK
dc.identifier.authorityCheah, KSE=rp00342en_HK
dc.identifier.authorityHuang, JD=rp00451en_HK
dc.description.naturepublished_or_final_versionen_HK
dc.identifier.doi10.1093/nar/gkg844en_HK
dc.identifier.pmid14602928-
dc.identifier.pmcidPMC275540en_HK
dc.identifier.scopuseid_2-s2.0-0344668833en_HK
dc.identifier.hkuros88445-
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0344668833&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume31en_HK
dc.identifier.issue22en_HK
dc.identifier.spage6674en_HK
dc.identifier.epage6687en_HK
dc.identifier.isiWOS:000186590600033-
dc.publisher.placeUnited Kingdomen_HK
dc.identifier.f10001016175-
dc.identifier.scopusauthoridLi, XT=8108660200en_HK
dc.identifier.scopusauthoridCostantino, N=6602994455en_HK
dc.identifier.scopusauthoridLu, LY=8686996700en_HK
dc.identifier.scopusauthoridLiu, DP=21934191400en_HK
dc.identifier.scopusauthoridWatt, RM=7102907536en_HK
dc.identifier.scopusauthoridCheah, KSE=35387746200en_HK
dc.identifier.scopusauthoridCourt, DL=7005765958en_HK
dc.identifier.scopusauthoridHuang, JD=8108660600en_HK

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