During bacterial mating, or conjugation, DNA is transferred from the donor cell to recipient cell through a mating pore. Mating is of great biomedical concern since it can transfer genes involved in disease and antibiotic resistance. The conjugative element ICEBs1 found in Bacillus subtilis encodes 25 putative proteins, one of which, ConE, is required for mating. . Although ConE's role in mating is somewhat unclear, ConE contains conserved ATPase motifs, indicating that it might provide the energy for DNA transfer. To explore ConE's ATPase activity in vitro, ConE was previously tagged with six histidines at the N terminus so that it could be purified by nickel chromatography. Here, we compared different wash buffers and quantities of resin to optimize the final protein product. Our new procedure provides higher yields of His6-ConE, which will enable us to explore its ATPase activity in vitro.

Purification of a bacterial mating protein

243rd National Meeting (2012)

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