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rdf:type |
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lifeskim:mentions |
umls-concept:C0012854,
umls-concept:C0205145,
umls-concept:C0439849,
umls-concept:C0442335,
umls-concept:C0444930,
umls-concept:C0445223,
umls-concept:C0547040,
umls-concept:C1515655,
umls-concept:C1552599,
umls-concept:C1704787,
umls-concept:C1881217
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pubmed:issue |
13
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pubmed:dateCreated |
2005-8-2
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pubmed:abstractText |
Contrary to several earlier reports, we find that cross-recombination between wild-type and the mutant loxP511 sites is <0.5% of that between two wild-type sites if Cre protein is expressed by phage P1 during an infection. The finding enabled us to develop a procedure to truncate DNA progressively from both ends of large genomic inserts flanked by these two loxP sites in pBACe3.6 and related vectors with transposons carrying either a wild-type or a loxP511 sequence. Newly constructed loxP511 transposons contained either a kanamycin resistance gene or no marker. Insert DNA ends in deletions were sequenced with primers unique to each transposon-end remaining after the respective recombination. End-sequencing 223 deletions confirmed that the low level of cross-recombination, observed between those sites during the P1 transductions, does not complicate the procedure: truncations from the unintended end of genomic inserts did not occur. Multiple BACs pooled together could also be processed in a single tube to make end-deletions. This deletion technology, utilizing the very minimal cross-recombination between the mutant and wild-type loxP sites of most BAC clones in the public domain and a heterologous one inserted as a transposon, should facilitate functionally mapping long-range gene regulatory sequences and help to isolate genes with defined functional boundaries in numerous projects including those of therapeutic interest.
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pubmed:grant |
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061933-10037821,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061933-10373322,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061933-10471489,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061933-10471751,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061933-10525404,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061933-10557311,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061933-10645956,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061933-10871388,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061933-10964509,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061933-10998271,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061933-11156622,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061933-11230172,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061933-11237328,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061933-11311165,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061933-11574153,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061933-11576551,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061933-12000846,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061933-12202778,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061933-12466304,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061933-12466850,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061933-12591158,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061933-12915490,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061933-1369729,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061933-15060022,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061933-15247347,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061933-1528894,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061933-15494454,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061933-15556570,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061933-2839833,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061933-3001054,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061933-3457367,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061933-7710783,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061933-7776370,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061933-7937090,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061933-8136839,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061933-9153322,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061933-9306400,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061933-9560239,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061933-9714735,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061933-9740665,
http://linkedlifedata.com/resource/pubmed/commentcorrection/16061933-9771703
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:issn |
1362-4962
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pubmed:author |
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pubmed:issnType |
Electronic
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pubmed:volume |
33
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
e118
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pubmed:dateRevised |
2009-11-18
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pubmed:meshHeading |
pubmed-meshheading:16061933-Bacteriophage P1,
pubmed-meshheading:16061933-Base Sequence,
pubmed-meshheading:16061933-Chromosomes, Artificial, Bacterial,
pubmed-meshheading:16061933-DNA,
pubmed-meshheading:16061933-DNA Primers,
pubmed-meshheading:16061933-DNA Transposable Elements,
pubmed-meshheading:16061933-Deoxyribonucleases, Type II Site-Specific,
pubmed-meshheading:16061933-Integrases,
pubmed-meshheading:16061933-Mutagenesis, Insertional,
pubmed-meshheading:16061933-Plasmids,
pubmed-meshheading:16061933-Recombination, Genetic,
pubmed-meshheading:16061933-Regulatory Sequences, Nucleic Acid,
pubmed-meshheading:16061933-Sequence Deletion,
pubmed-meshheading:16061933-Transduction, Genetic,
pubmed-meshheading:16061933-Viral Proteins
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pubmed:year |
2005
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pubmed:articleTitle |
Minimal cross-recombination between wild-type and loxP511 sites in vivo facilitates truncating both ends of large DNA inserts in pBACe3.6 and related vectors.
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