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Cloning ofmutHand identification of the gene product

Authors
Journal
Gene
0378-1119
Publisher
Elsevier
Publication Date
Volume
22
Identifiers
DOI: 10.1016/0378-1119(83)90109-9
Keywords
  • Recomhinant Dna
  • Imitator Gene
  • Recombinant Plasmids
  • Deletion Analysis: In Vitro Transcription-Translation
  • Maxicells
  • Promoter
  • Transducing Phage Lambda
Disciplines
  • Biology

Abstract

Abstract A specialized A transducing phage carrying the mutH gene and several deletion derivatives of this phage were characterised 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 HinddIII 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 M M r 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 M r 25000 protein.

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