Construction of a Viral Helicase Nanopore for Active DNA Unwinding and Transport

The molecular nanomotors play a key role in various biological process, such as DNA translocation in double-stranded DNA bacteriophage. Motors with the ATPase and helicase activity can unwind the double-strand DNA in the presence of ATP in vivo. Here we report the construction of a viral helicase and it displays the enzyme activity in vitro with the potential in material transport for nanomedicine. The helicase protein was purified by glutathione agarose and digestion GST tag and helicase protein by PreScission Protease, and further separated using AKTA purification system. The obtained viral helicase motor can unwind dsDNA both in vitro and in lipid bilayer membrane.