Cloning of Hydroxylamine Oxidoreductase from Nitrosomonas europaea into Shewanella oneidensis
Mentor 1
Dr. A. Andrew Pacheco
Location
Union Wisconsin Room
Start Date
28-4-2017 1:30 PM
End Date
28-4-2017 4:00 PM
Description
Hydroxylamine Oxidoreductase (henceforth HAO) is a trimeric protein found to contain 7 c-type hemes and one unique heme called P-460 for every monomer of the protein. HAO is a necessary part of the bacterium Nitrosomonas europaea's ability to participate in the nitrogen cycle. The function of HAO in N. europaea is to catalyze the reduction of hydroxylamine to nitrite. The primary goal of this project was to clone the HAO gene into the bacterium Shewenella oneidensis. Whereas N. europaea cultures take many weeks to grow a minimum usable optical density (OD), S. oneidensis cultures grow to a much higher OD overnight. This in turn means that substantially higher quantities of HAO can be purified from S. oneidensis than from N. europaea cultures, and they can be obtained more rapidly. This is an enormous advantage for experiments performed in the Pacheco group, which require large quantities of HAO. An important reason why S. oneidensis cells in particular were chosen is for their ability to readily produce c-type hemes. This poster will present the progress made to date in the attempt to clone HAO into S. oneidensis.
Cloning of Hydroxylamine Oxidoreductase from Nitrosomonas europaea into Shewanella oneidensis
Union Wisconsin Room
Hydroxylamine Oxidoreductase (henceforth HAO) is a trimeric protein found to contain 7 c-type hemes and one unique heme called P-460 for every monomer of the protein. HAO is a necessary part of the bacterium Nitrosomonas europaea's ability to participate in the nitrogen cycle. The function of HAO in N. europaea is to catalyze the reduction of hydroxylamine to nitrite. The primary goal of this project was to clone the HAO gene into the bacterium Shewenella oneidensis. Whereas N. europaea cultures take many weeks to grow a minimum usable optical density (OD), S. oneidensis cultures grow to a much higher OD overnight. This in turn means that substantially higher quantities of HAO can be purified from S. oneidensis than from N. europaea cultures, and they can be obtained more rapidly. This is an enormous advantage for experiments performed in the Pacheco group, which require large quantities of HAO. An important reason why S. oneidensis cells in particular were chosen is for their ability to readily produce c-type hemes. This poster will present the progress made to date in the attempt to clone HAO into S. oneidensis.
