The effect of Ce3+ on Pseudomonas Putida motility and biofilm formation
Emily Marie Breaux
Heme proteins are rarely studied in an academic laboratory setting due to their sensitivity and instability. Tt - HNOX is a signaling heme protein produced by thermophilic bacteria that is stable at room temperature and easy to purify. This study sought to optimize the Tt - HNOX expression, purification, and characterization process for the purpose of implementing the procedure in an educational lab. Various methods of BL21competent E. Coli (transformed to express the protein of interest) growth were tested and the optimal method was then used to repeatedly produce Tt - HNOX in high yield. Various methods of protein purification and characterization were then explored to find the most efficient ways to study the protein. For characterization, we analyzed Tt - HNOX in its unligated states and oxygen bound state. An optimal experimental method was found to consistently reproduce a high concentration of expressed protein that could then be accurately and thoroughly characterized.
Chemistry, Biochemistry and Molecular Biology