File Download
  Links for fulltext
     (May Require Subscription)
Supplementary

Article: Combination of overlapping bacterial artificial chromosomes by a two-step recombinogenic engineering method.

TitleCombination of overlapping bacterial artificial chromosomes by a two-step recombinogenic engineering method.
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. 15, p. e81 How to Cite?
AbstractRecombinogenic engineering or recombineering is a powerful new method to engineer DNA without the need for restriction enzymes or ligases. We report here a general method for using recombineering to combine overlapping bacterial artificial chromosomes (BACs) to build larger, unified BACs. In order to test the feasibility of using recombineering to combine two large DNA fragments (>20 kb), we constructed a unified BAC containing the full-length tyrosinase-related protein-1 (Tyrp-1) gene from two library-derived BACs, one containing the 5' regulatory elements and the other containing the 3' coding exons. This was achieved using a two-step homologous recombination method enabled by the bacteriophage lambda Red proteins. In the first step, retrieval, a large DNA fragment (approximately 22 kb) was retrieved from one of the original BACs. In the second step, recombination, the retrieved DNA fragment was inserted into the second original BAC to form the unified BAC containing all the desired Tyrp-1 sequence. To further demonstrate the general applicability of our approach, an additional DNA fragment (approximately 20 kb) was inserted into the unified BAC downstream of the coding region. This method should prove very useful for enabling BAC manipulation in a variety of scenarios.
Persistent Identifierhttp://hdl.handle.net/10722/48986
ISSN
2015 SCImago Journal Rankings: 7.458
PubMed Central ID
ISI Accession Number ID

 

DC FieldValueLanguage
dc.contributor.authorZhang, XMen_HK
dc.contributor.authorHuang, JDen_HK
dc.date.accessioned2008-06-12T06:31:25Z-
dc.date.available2008-06-12T06:31:25Z-
dc.date.issued2003en_HK
dc.identifier.citationNucleic Acids Research, 2003, v. 31 n. 15, p. e81en_HK
dc.identifier.issn1362-4962en_HK
dc.identifier.urihttp://hdl.handle.net/10722/48986-
dc.description.abstractRecombinogenic engineering or recombineering is a powerful new method to engineer DNA without the need for restriction enzymes or ligases. We report here a general method for using recombineering to combine overlapping bacterial artificial chromosomes (BACs) to build larger, unified BACs. In order to test the feasibility of using recombineering to combine two large DNA fragments (>20 kb), we constructed a unified BAC containing the full-length tyrosinase-related protein-1 (Tyrp-1) gene from two library-derived BACs, one containing the 5' regulatory elements and the other containing the 3' coding exons. This was achieved using a two-step homologous recombination method enabled by the bacteriophage lambda Red proteins. In the first step, retrieval, a large DNA fragment (approximately 22 kb) was retrieved from one of the original BACs. In the second step, recombination, the retrieved DNA fragment was inserted into the second original BAC to form the unified BAC containing all the desired Tyrp-1 sequence. To further demonstrate the general applicability of our approach, an additional DNA fragment (approximately 20 kb) was inserted into the unified BAC downstream of the coding region. This method should prove very useful for enabling BAC manipulation in a variety of scenarios.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.meshChromosomes, Artificial, Bacterialen_HK
dc.subject.meshGenetic Engineering - methodsen_HK
dc.subject.meshOxidoreductasesen_HK
dc.subject.meshProteins - geneticsen_HK
dc.subject.meshRecombination, Geneticen_HK
dc.titleCombination of overlapping bacterial artificial chromosomes by a two-step recombinogenic engineering method.en_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=0305-1048&volume=31&issue=15&spage=e81&epage=&date=2003&atitle=Combination+of+overlapping+bacterial+artificial+chromosomes+by+a+two-step+recombinogenic+engineering+methoden_HK
dc.identifier.emailHuang, JD:jdhuang@hkucc.hku.hken_HK
dc.identifier.authorityHuang, JD=rp00451en_HK
dc.description.naturepublished_or_final_versionen_HK
dc.identifier.doi10.1093/nar/gng081en_HK
dc.identifier.pmid12888533-
dc.identifier.pmcidPMC169968en_HK
dc.identifier.scopuseid_2-s2.0-0042662854en_HK
dc.identifier.hkuros87326-
dc.identifier.volume31en_HK
dc.identifier.issue15en_HK
dc.identifier.spagee81en_HK
dc.identifier.epagee81en_HK
dc.identifier.isiWOS:000184532900005-
dc.identifier.scopusauthoridZhang, XM=37092286900en_HK
dc.identifier.scopusauthoridHuang, JD=8108660600en_HK

Export via OAI-PMH Interface in XML Formats


OR


Export to Other Non-XML Formats