Shonda Campbell

Poster Presentation

Development of sensors to detect the frequency of translational misreading errors

High levels of recombinant protein expression have been known to place the hosts translational system under significant stress, causing an increase in mistranslation errors and reducing the yield of protein. To better understand how high levels of recombinant protein expression stresses the translation system, a dual reporter system was developed to measure the frequency of +1 frameshifting. The system contains an mcherry-GFP translational fusion, with a series of random frameshift sites designed such that the level of GFP expression is a measure of the frameshifting. To determine whether translational frameshifting increased when the cells were overexpressing a recombinant protein, the mCherry-GFP constructs were introduced into cells making high levels of the recombinant maltose binding protein (MBP; the stressor). We hypothesized that high levels of MBP would stress the translation system, and a higher level of translational frameshifting would be detected. Preliminary results show no significant difference between cells that only contain the reporter and cells that contain both the reporter and the stressor.