Linked Life Data

11239009

Source:http://linkedlifedata.com/resource/pubmed/id/11239009

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
6
pubmed:dateCreated
2001-3-12
pubmed:abstractText
The transformation-associated recombination (TAR) cloning technique allows selective and accurate isolation of chromosomal regions and genes from complex genomes. The technique is based on in vivo recombination between genomic DNA and a linearized vector containing homologous sequences, or hooks, to the gene of interest. The recombination occurs during transformation of yeast spheroplasts that results in the generation of a yeast artificial chromosome (YAC) containing the gene of interest. To further enhance and refine the TAR cloning technology, we determined the minimal size of a specific hook required for gene isolation utilizing the Tg.AC mouse transgene as a targeted region. For this purpose a set of vectors containing a B1 repeat hook and a Tg.AC-specific hook of variable sizes (from 20 to 800 bp) was constructed and checked for efficiency of transgene isolation by a radial TAR cloning. When vectors with a specific hook that was >/=60 bp were utilized, approximately 2% of transformants contained circular YACs with the Tg.AC transgene sequences. Efficiency of cloning dramatically decreased when the TAR vector contained a hook of 40 bp or less. Thus, the minimal length of a unique sequence required for gene isolation by TAR is approximately 60 bp. No transgene-positive YAC clones were detected when an ARS element was incorporated into a vector, demonstrating that the absence of a yeast origin of replication in a vector is a prerequisite for efficient gene isolation by TAR cloning.
pubmed:commentsCorrections
http://linkedlifedata.com/resource/pubmed/commentcorrection/11239009-10610723, http://linkedlifedata.com/resource/pubmed/commentcorrection/11239009-10915866, http://linkedlifedata.com/resource/pubmed/commentcorrection/11239009-11161779, http://linkedlifedata.com/resource/pubmed/commentcorrection/11239009-2251261, http://linkedlifedata.com/resource/pubmed/commentcorrection/11239009-2659436, http://linkedlifedata.com/resource/pubmed/commentcorrection/11239009-6449009, http://linkedlifedata.com/resource/pubmed/commentcorrection/11239009-7651842, http://linkedlifedata.com/resource/pubmed/commentcorrection/11239009-7789793, http://linkedlifedata.com/resource/pubmed/commentcorrection/11239009-8016135, http://linkedlifedata.com/resource/pubmed/commentcorrection/11239009-8321201, http://linkedlifedata.com/resource/pubmed/commentcorrection/11239009-8552668, http://linkedlifedata.com/resource/pubmed/commentcorrection/11239009-9016579, http://linkedlifedata.com/resource/pubmed/commentcorrection/11239009-9339466, http://linkedlifedata.com/resource/pubmed/commentcorrection/11239009-9457679, http://linkedlifedata.com/resource/pubmed/commentcorrection/11239009-9539761, http://linkedlifedata.com/resource/pubmed/commentcorrection/11239009-9585254
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Mar
pubmed:issn
1362-4962
pubmed:author
pubmed:issnType
Electronic
pubmed:day
15
pubmed:volume
29
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
E32
pubmed:dateRevised
2009-11-18
pubmed:meshHeading
pubmed:year
2001
pubmed:articleTitle
Defining the minimal length of sequence homology required for selective gene isolation by TAR cloning.
pubmed:affiliation
Laboratory of Molecular Genetics and Laboratory of Molecular Carcinogenesis, National Institute of Environmental Health Sciences, Research Triangle Park, NC 27709, USA.
pubmed:publicationType
Journal Article