Genetic and physical maps of Klebsiella aerogenes genes for histidine utilization ( hut )

Abstract

Deletion derivatives of the hut -containing plasmid pCB101 were tested against point mutants defective in individual genes of the histidine utilization ( hut ) operons using a complementation/recombination assay. Location of the genes of the right operon, hutU and hutH , was confirmed by direct assay of the gene products, urocanase and histidase; location of the repressor gene was identified by measuring the ability of the plasmid-carried genes to repress the formation of histidase from a chromosomal location. The analysis of eight deletion plasmids unambiguously confirms the map order of the hut genes as hutI-G-C-U-H , and demonstrates that, in Klebsiella aerogenes , the hutU and hutH genes are transcribed from their own promoter. In addition, the genetic map of hut can be aligned with the restriction map of the hut DNA in plasmid pCB101. One of the deletion plasmids studied apparently encodes a defective histidase subunit that is trans-dominant to active histidase. Another deletion, which completely removes the left operon, hutIG , allows high level expression of the hutUH operon and thus overproduction of a toxic intermediate.