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Cloning and characterization of a DNA repair gene inHaemophilus influenzae
Authors:Rohinee P. Kanade  N. K. Notani
Affiliation:(1) Molecular Biology and Agriculture Division, Bhabha Atomic Research Centre, 400 085 Bombay, India
Abstract:Using a high-efficiency DNA cloning vector pJ1–8, a DNA repair geneuvr1 has been self-cloned in bacteriumHaemophilus influenzae. Chimeric plasmid pKuvrl, carrying wild type allele ofuvr1 gene and flanking DNA sequences, specifically complements auvr1 gene mutation in the bacterial chromosome. Auvr1} mutation could be transferred from chromosome byin vivo recombination to pKuvr1 and isolated and designated as plasmid pKuvrl. Plasmid pKuvrl carries a 11.3 kb chromosomal DNA insert which was scanned for the presence of any other DNA repair genes by a novel method of directed mutagenesis. Preliminary analysis of the 3 new mutants isolated by this method supports the notion that the insert contains more than one gene concerned with ultraviolet radiation-sensitivity.
Keywords:Haemophilus influenzae   self-cloning   uvr1 gene  directed mutagenesis
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