Environmental Science and Management (ESM)
Bayer School of Natural and Environmental Sciences
molybdoenzyme, over expression
Periplasmic nitrate reductase from Sulfurospirillum barnesii is representative of the distinct branch of epsilonproteobacteria periplasmic nitrate reductases. The gene encoding the active subunit of S. barnesii's periplasmic nitrate reductase (napA) was successfully cloned into the vector pET101/D/TOPO. Constructs were made both with and without the sequence coding for the periplasmic signal sequence. Preliminary expression and purification experiments were performed with limited success. The DNA encoding the periplasmic signal sequence proved necessary for expression. Even with the signal sequence coding region, protein expression was limited. The low levels of expression appeared due to product degradation during cell growth. Purification was incomplete, with both low recovery levels and significant contamination.
Bird, L. (2007). Cloning and Heterologous Expression of the napA Gene from Sulfurospirillum barnesii (Master's thesis, Duquesne University). Retrieved from https://dsc.duq.edu/etd/319