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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
17
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pubmed:dateCreated |
1983-1-19
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pubmed:abstractText |
Deletion loop mutagenesis is a new, general method for site-directed mutagenesis that allows point mutations to the introduced within a sequence of DNA defined by a previously isolated deletion mutant. Wild type and deletion mutant DNA are cloned into a bacterial plasmid and each is cleaved with a different single cut restriction enzyme. Heteroduplexes are formed between the two DNAs to produce circular molecules containing a nick in each strand and a single-stranded deletion loop. The deletion loops are mutagenised using sodium bisulphite and the DNA transfected directly into a uracil repair deficient strain of Escherichia coli. Up to half of the resultant clones contain DNA produced by replication of the wild-type length strand and bear mutations exclusively within the target area. An example is given in which a deletion mutant lacking 21 nucleotides from the region coding for SV40 large-T was used. Eight of the possible nine target cytosine residues were mutagenised. The method described is specific, efficient and simple.
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/6292832-157836,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6292832-178902,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6292832-209457,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6292832-226361,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6292832-2948,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6292832-350837,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6292832-383576,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6292832-388356,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6292832-392601,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6292832-6246368,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6292832-6254078,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6292832-6259538,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6292832-6259539,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6292832-6276753,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6292832-6277737,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6292832-6279018,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6292832-6281730,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6292832-6282692,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6292832-6329679,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6292832-6460194,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6292832-681366,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6292832-7041119,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6292832-712841,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6292832-7273136,
http://linkedlifedata.com/resource/pubmed/commentcorrection/6292832-769308
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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:month |
Sep
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pubmed:issn |
0305-1048
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:day |
11
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pubmed:volume |
10
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
5161-71
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pubmed:dateRevised |
2009-11-18
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pubmed:meshHeading |
pubmed-meshheading:6292832-Base Sequence,
pubmed-meshheading:6292832-Chromosome Deletion,
pubmed-meshheading:6292832-DNA, Bacterial,
pubmed-meshheading:6292832-DNA Restriction Enzymes,
pubmed-meshheading:6292832-Escherichia coli,
pubmed-meshheading:6292832-Mutation,
pubmed-meshheading:6292832-Nucleic Acid Hybridization,
pubmed-meshheading:6292832-Plasmids,
pubmed-meshheading:6292832-Sulfites,
pubmed-meshheading:6292832-Transfection
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pubmed:year |
1982
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pubmed:articleTitle |
Deletion loop mutagenesis: a novel method for the construction of point mutations using deletion mutants.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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