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Molecular cloning and characterization of a genetic region from Serratia marcescens involved in DNA repair
Author(s) -
Murphy K. E.,
Braymer H. D.
Publication year - 1989
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1111/j.1365-2958.1989.tb01814.x
Subject(s) - biology , serratia marcescens , plasmid , complementation , molecular cloning , escherichia coli , dna , microbiology and biotechnology , mutant , genetics , gene , peptide sequence
Summary We report here the molecular isolation of a DNA fragment which encodes Tag‐like activity from the Gram‐negative bacterium Serratia marcescens. A recombinant plasmid encoding Tag‐like activity was isolated from a S. marcescens plasmid gene library by complementation of an Escherichia coli tag mutant, which is deficient in 3‐methyladenine DNA glycosylase I. The clone complements E. coli tag, recA, alkA , but not alkB , mutants for resistance to the DNA‐damaging agent methyl methanesulphonate (MMS). The coding region of the Tag activity, initially isolated on a 6.5kb BamHI fragment, was defined to a 1.8kb Bgl II‐ Smal fragment. Labelling of plasmid‐encoded proteins using maxicells revealed that the 1.8kb fragment encodes two proteins of molecular weights 42000 and 16000. Data presented here suggest that the cloned fragment encodes a DNA repair protein(s) that has similar activity to the 3‐methyladenine DNA glycosylase I of E. coli.