Preparation of Elongation Factor G for Electron Paramagnetic Resonance Analysis

In this research, the determination of the structure of Escherichia Coli Elongation Factor G (EFG) by Electron Paramagnetic Resonance (EPR) analysis was attempted. Expression of protein was performed using both BL21 and LEMO21 cells in both LB broth and minimal media. However, the expression of EFG was unsuccessful in all trials. The integrity of the plasmid containing the EFG gene was also analyzed by sequencing and restriction digest. It was found through sequencing that all the plasmids had a section of the EFG gene and plasmid DNA cut out, meaning that they would not create viable EFG protein. These results suggest that the EFG plasmids taken from previous years of research are unusable, and that they must be remade and the project restarted.