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TARGETED MUTAGENESIS OF M13 DNA INDUCED BY PHOTOSENSITIZED REACTION
Author(s) -
Piette Jacques,
Van De Vorst Albert
Publication year - 1984
Publication title -
photochemistry and photobiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.818
H-Index - 131
eISSN - 1751-1097
pISSN - 0031-8655
DOI - 10.1111/j.1751-1097.1984.tb05352.x
Subject(s) - proflavine , mutagenesis , dna , gene , microbiology and biotechnology , base pair , transfection , biology , chemistry , genetics , mutation
—Inactivation and mutagenesis of M13 mp10 DNA either on a double‐stranded or a single‐stranded form have been induced after photosensitization mediated by proflavine. Both the rate of inactivation and the frequency of the induction of mutations in the Lac gene are at least ten times more important when the double‐stranded DNA has been used as substrate for the photosensitization reaction. Promazine has also been able to promote photomutagenesis in the Lac gene but less efficiently than proflavine. Numerous clones coming from E. cali K12 JM103 transfection by photosensitized single‐ or double‐stranded M13 mp10 DNA have been purified and amplified. The DNA of each individual clone has been extracted and sequenced in the Lac gene using two different primers. An important hot spot for photosensitized mutagenesis has been localized at position ‐65 from the start of the Lac gene transcription and corresponds to a transition pattern G → A. Other minor base sequence modifications have also been recorded in the Lac gene.