Targeted mutagenesisin vitro: lacrepressor mutations generated-using AMV reverse transcriptase and dBrUTP
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Targeted mutagenesis in vitro: lac repressor mutations generated using AMV reverse transcriptase and dBrUTP.
We have cloned the gene for the lac operon repressor (lacI) of Escherichia coli into the M13 related phage f1. Mutagenesis of the lacI gene was performed in vitro by filling dsDNA molecules gapped over the lacI gene with Avian Myeloblastosis Virus (AMV) reverse transcriptase. LacI mutants are found at a frequency of 1 in 10(4) using a genetic screen in vivo. For two-thirds of the 60 mutants, le...
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ژورنال
عنوان ژورنال: Nucleic Acids Research
سال: 1984
ISSN: 0305-1048,1362-4962
DOI: 10.1093/nar/12.10.4139