Cyanobacteria possess an inducible mechanism which enables them to concentrate inorganic carbon (Ci) within the cells. An inactivation library was used to raise the high-CO2-requiring mutant of Synechococcus PCC 7942, IL-2, impaired in HCO3- transport. Analysis of the relevant genomic DNA detected several modifications, probably due to the single crossover recombination, leading to inactivation of ORF467 (designated ictB) in IL-2. IctB contains 10 trans-membrane regions and is homologous to several transport-related proteins from various organisms. Kinetic analyses of HCO3- uptake in the wild type and IL-2 suggested the presence of two or three HCO3- carriers exhibiting different affinities to HCO3-.