Cloning and Expression of Glutamate Dehydrogenase A (gdhA) from the Cockroach Endosymbiont Blattabacterium Spp.
PublisherThe University of Arizona.
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AbstractCockroaches (Dictyoptera:Blattaria) have a unique ability to store excess nitrogen as uric acid in their fat bodies. It has been suggested that cockroach bacterial endosymbionts may be responsible for using stored nitrogen in fat bodies and turning it into usable nitrogen (e.g. amino acids). Recent sequence data has suggested that the cockroach endosymbiont Blattabacterium spp. could have nitrogen recycling ability through urease (ureA) and glutamate dehydrogenase (gdhA). To determine whether Blattabacterium gdhA is able to use ammonia, an assay was developed to first express the protein in E. coli and then observe ammonia activity through spectroscopy as well as ammonia activity of the purified protein. Blattabacterium gdhA was cloned into E. coli expression vectors, and the protein expression levels were assessed using SDS protein gels and Bradford?s Reagent assays. Detectable expression levels were not found in the SDS gel or Bradford Reagent, suggesting that the gdhA protein may be expressed at levels difficult to detect with the current assays. This work invites further exploration of this exceptional system through improved techniques and revised experimental design.
Degree ProgramHonors College
Biology - Biomedical Sciences