Iterative selection from a Salmonella typhimurium cosmid library can lead to the isolation of an atypically small plasmid

Abstract

We have constructed a DNA library from a virulent Salmonella typhimurium strain, in an avirulent strain. The process of selecting the components of interest from the library involved iterative growth in liquid culture. This resulted, after four cycles, in the culture becoming homogeneous for a single plasmid, which was much smaller than the average size for the library. We have identified the larger precursor of this plasmid which has two regions of sufficient homology to allow recombination resulting in the formation of the small plasmid. S. typhimurium carrying the small plasmid have a smaller genetic burden than other members of the library and survive better in spent culture medium, facilitating selection on repeated subculture. Such rapid adventitious selection has important implications for isolation of clones of interest from genomic libraries. z 1999 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.