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  • Title: Cloning of mutH and identification of the gene product.
    Author: Grafstrom RH, Hoess RH.
    Journal: Gene; 1983; 22(2-3):245-53. PubMed ID: 6307825.
    Abstract:
    A specialized lambda transducing phage carrying the mutH gene and several deletion derivatives of this phage were characterized by restriction enzyme analysis. This analysis localized the mutH gene to a small region of bacterial DNA on the transducing phage and facilitated the subsequent cloning of this gene into the multicopy plasmid pBR322. The mutH gene is contained entirely on a 1.5-kb HindIII fragment as judged by the ability of plasmids carrying this fragment to complement mutH- alleles on the bacterial chromosome. Using recombinant plasmids containing the 1.5-kb HindIII fragment, we identified an Mr 25000 protein as the product of the mutH gene in an in vitro transcription-translation system as well as in maxicells. Various deletion derivatives of the mutH-containing plasmids that exhibit a Mut- phenotype also have lost the Mr 25000 protein.
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