SUMMARY: The enzymes involved in gluconate utilization in seemed to be gluconate permease and gluconate kinase. Several mutants unable to grow on gluconate were isolated. The mutations they harboured () were clustered between and on the chromosome (a tentative map order of and was obtained). The mutation seemed to be located within the structural gene of the kinase, and the and mutations seemed to be within that of the permease. An RI fragment (4·5 MDal) containing an intact gluconate () operon consisting of these two structural genes was cloned in phage ϕ 105 by prophage transformation and was mapped physically. The physical location of the mutations coincided with their order on the genetic map. The dIII-A fragment (2·4 MDal), which corrects all the mutations, was subcloned in plasmid pC194. The fragment contained the structural genes for the gluconate permease and kinase, but not the regulatory region of the gluconate operon.


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