Cloning and Expression of Glutamate Dehydrogenase A (gdhA) from the Cockroach Endosymbiont Blattabacterium Spp.

Persistent Link:
http://hdl.handle.net/10150/146916
Title:
Cloning and Expression of Glutamate Dehydrogenase A (gdhA) from the Cockroach Endosymbiont Blattabacterium Spp.
Author:
Wong, Ryan Christopher
Issue Date:
May-2010
Publisher:
The University of Arizona.
Rights:
Copyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author.
Abstract:
Cockroaches (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.
Type:
text; Electronic Thesis
Degree Name:
B.S.
Degree Level:
bachelors
Degree Program:
Honors College; Biology - Biomedical Sciences
Degree Grantor:
University of Arizona

Full metadata record

DC FieldValue Language
dc.language.isoenen_US
dc.titleCloning and Expression of Glutamate Dehydrogenase A (gdhA) from the Cockroach Endosymbiont Blattabacterium Spp.en_US
dc.creatorWong, Ryan Christopheren_US
dc.contributor.authorWong, Ryan Christopheren_US
dc.date.issued2010-05-
dc.publisherThe University of Arizona.en_US
dc.rightsCopyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author.en_US
dc.description.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.en_US
dc.typetexten_US
dc.typeElectronic Thesisen_US
thesis.degree.nameB.S.en_US
thesis.degree.levelbachelorsen_US
thesis.degree.disciplineHonors Collegeen_US
thesis.degree.disciplineBiology - Biomedical Sciencesen_US
thesis.degree.grantorUniversity of Arizonaen_US
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