Regulation of toxinogenesis in Corynebacterium diphtheriae. II. Genetic mapping of a tox regulatory mutation in bacteriophage beta.

The structural gene for diphtherial toxin is present in corynebacteriophage beta. Previous studies located several point mutations within the tox locus and determined the orientation of transcription of the toxin structural gene. The production of maximal quantities of toxin by Corynebacterium diphtheriae C7(beta) occurs only when the bacteria are iron deficient. Mutations in phage beta can affect this control of toxin production by iron. The tox-201 mutation regulates expression of the toxin structural gene in a cis-dominant manner and permits large amounts of toxin to be made under high-iron conditions when phage beta tox-201 infects C. diphtheriae C7. In this study tox-201 was found to be closely linked to the structural gene for toxin. We performed a series of multifactor matings to determine the relative positions of tox-201 and several point mutations within the toxin structural gene. The order of these markers on the vegetative genetic map of phage beta was tox-201-tox-4-tox-2-tox-30. These findings establish that the tox-201 regulatory site is closely linked to the end of the toxin structural gene corresponding to the origin of transcription. This location is consistent with our hypothesis that tox-201 defines a cis-dominant regulatory element, such as an operator, promoter, or attenuator, involved in control of toxinogenesis in C. diphtheriae C7(beta).