We have previously shown that a mutation (groPC259) in the E. coli dnaJ gene renders the cell inviable at high temperatures and arrests bacteriophage lambda DNA replication at all temperatures (Sunshine et al., 1977). We have isolated lambda dnaJ+ transducing phages both by in vitro cloning and by abnormal excision of a lambda dnaK transducing phage integrated near the dnaJ locus. The dnaJ gene product has been identified on SDS polyacrylamide gels after infection of UV-irradiated E. coli cells by lambda dnaJ+ derivative phages. It is a polypeptide chain with an apparent molecular weight of 37,000-daltons. This has been verified by the fact that a transducing phage carrying an amber mutation in the dnaJ gene fails to induce the synthesis of the 37,000-dalton polypeptide chain upon infection of sup+ bacteria, but does so upon infection of supF or supD bacteria.