A fine structure map of spontaneous and induced mutations in the lambda repressor gene, including insertions of IS elements

Abstract

Mutations at over 70 sites in the cI gene have been mapped by 4-factor crosses and assigned precise or approximate positions in the DNA sequence. 16 of 25 spontaneous mutations were insertions of IS1, IS3 or IS5 into AT-rich regions of cI. The 5-methylcytosine in the sequence Cm5CAGG is a hot spot for spontaneous cI amber mutations. Recombination frequencies between mutations were proportional to distance with the exception of amber mutations at 4 sites, including the host spot for spontaneous mutations. Mutations with a given phenotype are clustered on the genetic map. No missense mutations affecting repressor activity were found in the central one-third of cI, but 5 of 6 ind- mutations were located in this region. The amino-terminal third of the gene contains the sites of most trans-dominant cI- mutations, and of all ts mutations that result in repressors that are reversibly inactivated at high temperatures.