Extant Escherichia coli K12 strains are phenotypically rough, their li
popolysaccharide having a complete core structure, but no O antigen. W
e used DNA hybridization and DNA sequencing to show that the rough phe
notype of this strain is due to the presence of one of two independent
mutations in the rfb gene cluster. The rfb-50 mutation, consisting of
an IS5 insertion at the downstream end of rfb, is present in strain E
MG2, which is representative of most K12 derivatives. The rfb-51 mutat
ion is a deletion at the upstream end of rfb, and was found in strain
WG1. A gene cloned from strain WG1 could complement the rfb-50 mutatio
n in strain EMG2, and the complemented strain produced O antigen which
was typed as O16 with cross reaction to O17.