Purification and characterization of DNA dependent RNA polymerase from Staphylococcus aureus

DNA dependent RNA polymerase from exponentially growing Staphylococcus aureus cells was purified. An SDS-polyacrylamide gel analysis of the most purified preparation revealed that it consists of beta, beta', alpha, and sigma with apparent molecular masses of 151, 147, 42, and 55 kDa, respectively. The sigma subunit cross reacted with a polyclonal antibody against Bacillus subtilis sigma 43. The cross reacting peptide co-migrated with the B. subtilis sigma 43 subunit. The implications of these results are discussed. Promoter specific in vitro run-off transcripts were obtained using the purified enzyme preparation. Specific conditions for the polymerization reaction are defined.