Creative Commons License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License.
Using Polymerase Chain Reaction to Detect nifH and vnfDGK in a Novel Acidophilic Microbial Community
Acidophiles are widely known for their ability to exploit harsh environments, typically consisting of high temperatures, dissolved metal concentrations, and acidic conditions. Consequently, different acidophilic species are employed industrially in applications such as biomining and bioleaching. However, their full potential in bioremediation, and in industrial practices, has yet to be fully explored because many of these acidophiles are difficult to culture in laboratory conditions or have yet to be properly identified. This investigation focuses on a novel community sampled from the Richmond Mine site in Iron Mountain, California, consisting of a filamentous fungus and prokaryotes. This consortium can be cultured in laboratory conditions without the presence of organic nitrogen sources. Previous data has also revealed that the community has a nutritional requirement for vanadium ions. Therefore, I predicted that this bacterium is able to fix atmospheric nitrogen via the nitrogenase enzyme, and might do so using a rare variant that requires vanadium as a cofactor. My investigation will employ PCR amplification, ligation, and transformation as well as bioinformatics tools in attempts to isolate conserved nifH and vnfDGK gene fragments, that are found in standard nitrogenases and vanadium-dependent nitrogenases, respectively. Results from this investigation can be used not only to further understand the dynamics of the community found in these mine waters, but also to shed light on the process of nitrogen fixation in such a harsh environment.
Flowers, Samantha L., "Using Polymerase Chain Reaction to Detect nifH and vnfDGK in a Novel Acidophilic Microbial Community" (2011). Undergraduate Honors Theses. 38.