Gene Recombination and Segregation of Resistance Factor R in Escherichia coli

Abstract

Hashimoto, Hajime (Osaka University, Osaka, Japan), and Yukinori Hirota . Gene recombination and segregation of resistance factor R in Escherichia coli . J. Bacteriol. 91: 51–62. 1966.—Independent chloramphenicol-sensitive (CM s ) mutants of the drug-resistance factor R were isolated. Introduction of two different R factor CM s mutants into a single bacterium, by conjugation or transduction, gave chloramphenicol-resistant (CM r ) colonies when such strains were plated on a medium containing chloramphenicol (Cm). These CM r colonies resulted from recombination between two R factors contained within the same cell. Most of the CM r colonies were heterogeneous, and segregation of drug-resistance markers was observed among the progeny. Segregated bacteria which still carried the recombinant R factor were stable for resistance to Cm as well as for other markers of R. All the markers of recombinant R factors were cotransducible with high coincidence and at the same frequency as wild-type R. Sensitive mutants of R which had lost all the resistance markers of the R factor were found also. A mutation of R, referred to as SMA, which was sensitive to streptomycin and sulfanilamide, was capable of reverting to resistance to both of these drugs simultaneously. The sensitive alleles for SMA, CM, and TC were shown to be recessive to the resistance alleles. Mutants of R having multisite mutations or deletions in the CM gene were isolated and used to analyze the pattern of linked segregation of unselected markers of the recombinant R factor. The drug resistance factor R was shown to have two linkage groups, CM—SMA and TC— m .