Challenges in visualizing RecA-DNA interaction in chemiluminescent electrophoretic mobility shift assays

• In Event: Chemistry and Biochemistry Oral Session I

Abstract

A high-end DSLR camera was employed for analysis of protein-DNA interaction using chemiluminescent electrophoretic mobility shift assays (EMSAs) in order to study binding affinity of DNA repair protein, RecA, and selected Mycobacterium tuberculosis oligomers. This alternative method for EMSA visualization offers a low-cost benefit over using a phosphorimager with radioactively labeled DNA. Mini-format chemiluminescent, fluorescent, and large-format chemiluminescent EMSAs were used to determine percent RecA binding. Decrease in free DNA signal indicates RecA did bind to DNA, though a bound DNA band was not resolved in any of the three types of EMSAs. Future research will involve further examination of this protein-DNA interaction in order to determine why the complex is not migrating into the EMSAs.

Author

• Jessica Rose Kenneson, Wayland Baptist University