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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
5
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pubmed:dateCreated |
1982-6-21
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pubmed:databankReference |
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pubmed:abstractText |
A method for highly efficient segment-specific mutagenesis is described. The method uses as target for sodium bisulfite mutagenesis the DNA single strands of a DNA restriction fragment that had been separated by cloning into base-complementary regions of a pair of phage fd vectors. After repair synthesis in vitro, the mutagenized DNA fragment is recovered by cloning into a nonmutated plasmid vector and analyzed for sequence and by functional tests. By using this method, the nucleotide sequence of a 109-base pair restriction fragment containing the lac promoter/operator from Escherichia coli was extensively modified. More than 90% of the 235 isolates obtained showed a change in phenotype; all of 22 analyzed for their nucleotide sequence were found to carry multiple C leads to T point mutations in up to 60% of the possible target positions. Nevertheless, few isolates showed major changes in promoter activity relative to the nonmutated promoter element, which indicates a high degree of flexibility in the promoter sequence.
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/7041119-1069185,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7041119-10793721,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7041119-118083,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7041119-157836,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7041119-209457,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7041119-364474,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7041119-371668,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7041119-383387,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7041119-384366,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7041119-416358,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7041119-4310285,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7041119-4444051,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7041119-4555447,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7041119-4587255,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7041119-4598642,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7041119-4736082,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7041119-5449768,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7041119-6101056,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7041119-6245337,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7041119-6246368,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7041119-6248238,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7041119-6253889,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7041119-6254078,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7041119-6259615,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7041119-712841,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7041119-769308,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7041119-881736,
http://linkedlifedata.com/resource/pubmed/commentcorrection/7041119-94251
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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 |
Mar
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pubmed:issn |
0027-8424
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:volume |
79
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
1408-12
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pubmed:dateRevised |
2009-11-18
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pubmed:meshHeading |
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pubmed:year |
1982
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pubmed:articleTitle |
Segment-specific mutagenesis: extensive mutagenesis of a lac promoter/operator element.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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