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NAD+-dependent DNA ligases of Mycobacterium tuberculosis and Streptomyces coelicolor

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WILEY
DOI: 10.1002/prot.10361

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DNA end-joining; DNA repair; ligation; mycobacteria; streptomycetes

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Sequencing of the genomes of Mycobacterium tuberculosis H37Rv and Streptomyces coelicolor A3(2) identified putative genes for an NAD(+)-dependent DNA ligase. We have cloned both open reading frames and overexpressed the protein products in Escherichia coli. In vitro biochemical assays confirm that each of these proteins encodes a functional DNA ligase that uses NAD(+) as its cofactor. Expression of either protein is able to complement E. coli GR501, which carries a temperature-sensitive mutation in ligA. Thus, in vitro and in vivo analyses confirm predictions that ligA genes from M. tuberculosis and S. coelicolor are NAD(+)-dependent DNA ligases. Proteins 2003;51:321-326. (C) 2003 Wiley-Liss, Inc.

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