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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
2
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pubmed:dateCreated |
1989-3-21
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pubmed:abstractText |
In the polymerase chain reaction (PCR) the specific amplification of a small segment of DNA within a complex DNA sample is effected by repeated cycles of DNA denaturation and enzymatic synthesis primed by two oligonucleotides complementary to regions within opposite strands of the DNA. In this report a simple and efficient method is described in which PCR methodology is used to introduce specific mutations into a double stranded DNA molecule. In this procedure a supercoiled plasmid DNA serves as template for a PCR in which a primer bearing the mutated sequence is incorporated into the amplified product. The presence of convenient restriction sites in the mutagenic primer and in the amplified DNA permit direct replacement of a wild type DNA segment with the mutated segment by treating the PCR mixture with the appropriate restriction endonucleases followed by DNA ligase. Using this procedure, a single amino acid replacement, a 16 amino acid deletion and a replacement of four amino acids with a twelve amino acid segment from another membrane protein were introduced into the amino terminal signal segment of rat hepatic cytochrome P450b (P450IIB1).
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pubmed:grant |
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/2915928-2448875,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2915928-271968,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2915928-2847780,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2915928-2881260,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2915928-2895676,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2915928-2989270,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2915928-2999980,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2915928-3002241,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2915928-3045756,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2915928-3047140,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2915928-3110950,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2915928-3137560,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2915928-3267215,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2915928-3278322,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2915928-3323813,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2915928-3336784,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2915928-3344434,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2915928-3431465,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2915928-3461561,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2915928-3587348,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2915928-3755079,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2915928-3881765,
http://linkedlifedata.com/resource/pubmed/commentcorrection/2915928-6096830
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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 |
Jan
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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 |
25
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pubmed:volume |
17
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
723-33
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pubmed:dateRevised |
2009-11-18
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pubmed:meshHeading |
pubmed-meshheading:2915928-Animals,
pubmed-meshheading:2915928-Base Sequence,
pubmed-meshheading:2915928-Chimera,
pubmed-meshheading:2915928-Chromosome Deletion,
pubmed-meshheading:2915928-Cloning, Molecular,
pubmed-meshheading:2915928-Cytochrome P-450 Enzyme System,
pubmed-meshheading:2915928-DNA Mutational Analysis,
pubmed-meshheading:2915928-DNA-Directed DNA Polymerase,
pubmed-meshheading:2915928-Gene Amplification,
pubmed-meshheading:2915928-Gene Conversion,
pubmed-meshheading:2915928-Genes, Synthetic,
pubmed-meshheading:2915928-Oligonucleotide Probes,
pubmed-meshheading:2915928-Rats,
pubmed-meshheading:2915928-Taq Polymerase
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pubmed:year |
1989
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pubmed:articleTitle |
Construction of mutant and chimeric genes using the polymerase chain reaction.
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pubmed:affiliation |
Department of Cell Biology, New York University School of Medicine 10016.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.
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