Production of Reverse Transcriptase and DNA Polymerase in Bacterial Expression Systems.

DNA amplification and reverse transcription enzymes have proven to be invaluable in fast and reliable diagnostics and research applications because of their processivity, specificity, and robustness. Our study focused on the production of mutant Taq DNA polymerase and mutant M-MLV reverse transcriptase in the expression hosts and under various expression conditions. We also examined nonspecific extracellular production in . Intracellularly, M-MLV was produced in at the level of 11% of the total cell proteins (TCPs) compared with 16% of TCPs in . We obtained a soluble protein that accounted for 11% of the enzyme produced in and 22% of the enzyme produced in . Taq pol was produced intracellularly in at the level of 30% of TCPs compared with 26% of TCPs in . However, Taq pol was almost non-soluble in , whereas in , we obtained a soluble protein that accounted for 23% of the produced enzyme. We detected substantial extracellular production of Taq pol. Thus, is a suitable alternative host with the potential for production of recombinant proteins.