Two proteins, barnase, the extracellular ribonuclease of Bacillus amyloliquefaciens, and barstar, its intracellular inhibitor, are used as a model system for the study of protien folding and protein-protein interactions. Barnase is one of an homologous group of ribonucleases occurring in both prokaryotes and eukaryotes. Amino acid sequence determination and X-ray structure characterization are being carried out along with theromydynamic studies of folding reactions. A peptide complementation system for barnase has been developed which will be used to investigate the roles of specific residues in both folding and enzymatic activity. Recombinant DNA techniques are being applied to the project with three major aims: 1) to facilitate production, 2) to examine the structural and control sequences of the genes and 3) to tailor specifically designed modifications in the sequences to test theories of protein folding. A technique of transposon mutagenesis has been adapted to B. amyloliquefaciens. This permits the identification and cloning of any gene for which there is a functional test. A gene controlling the synthesis of barnase has been found and, with its inserted transposon, cloned. This DNA has been partially sequenced and identified us the barnase structural gene.