A BAC library from common bean has been used in order to isolate the entire multigene Bowman-Birk serine protease inhibitor family and to study its genome organization. Using a previously isolated trypsin/chymotrypsin inhibitor nucleotide sequence as probe, two positive BAC clones were identified. The P2B8 BAC clone, of about 135 kbp and containing the complete BBI family, was chosen and partially sequenced. Our results confirm that a small multigene family codes for three double-headed inhibitors named: tc-BBI-1, tc-BBI-2 and et-BBI. They contain the binding loop trypsin/chymotrypsin (tc-BBI-1 and tc-BBI-2) and the elastase/trypsin one (et-BBI), respectively. Genes coding for tc-BBI-1 and et-BBI, were found to be very close to each other and arranged in a head to head fashion. Southern blot hybridisation on genomic DNA digested with PstI enzyme suggests that all three genes are present in a fragment of 19 kbp. Northern blot analyses on RNA isolated from various common bean organs showed that the expression of tc-BBI-1 and et-BBI was restricted to the developing cotyledons.
