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CHARACTERIZATION OF CYTOCHROME C PEROXIDASE OF
MARINOBACTER AQUAEOLEI
by
Jamie Lynn Waite
________________________________________________________________________
A Thesis Presented to the
FACULTY OF THE USC GRADUATE SCHOOL
UNIVERSITY OF SOUTHERN CALIFORNIA
In Partial Fulfillment of the
Requirements for the Degree
MASTER OF SCIENCE
(BIOCHEMISTRY AND MOLECULAR BIOLOGY)
May 2012
Copyright 2012 Jamie Lynn Waite
Object Description
| Title | Characterization of cytochrome c peroxidase of Marinobacter aquaeolei |
| Author | Waite, Jamie Lynn |
| Author email | jamielynn.waite@gmail.com;jamielynn.waite@gmail.com |
| Degree | Master of Science |
| Document type | Thesis |
| Degree program | Biochemistry and Molecular Biology |
| School | Keck School of Medicine |
| Date defended/completed | 2012-01-20 |
| Date submitted | 2012-04-13 |
| Date approved | 2012-04-13 |
| Restricted until | 2012-04-13 |
| Date published | 2012-04-13 |
| Advisor (committee chair) | Tokes, Zoltan |
| Advisor (committee member) |
Edwards, Katrina J. Hacia, Joseph |
| Abstract | The molecular biology and biochemistry of biological iron oxidation at neutral pH is poorly understood. In part this is due to the fact that most chemolithoautotrophic iron oxidizing bacteria are genetically intractable and very difficult to cultivate and manipulate in the laboratory. The Marinobacter are ubiquitous marine bacteria that occur in hydrothermal settings, including hydrothermal plumes, chimneys, and seafloor exposed basalts. Many are capable of iron oxidation and can also be readily cultivated and studied in the laboratory. We hypothesize Marinobacter aquaeolei, a lithoheterotrophic iron oxidizing bacterium, utilizes the gene cytochrome c peroxidase (CCP) for Fe-oxidation during lithoheterotrophic growth. We have characterized heme protein activity during Fe-oxidation via heme staining followed by searching NCBI for genes of interest. Our preliminary research led us to cytochrome c peroxidase, a small heme protein. We synthetically mutated CCP, ligated the mutant into pJP5603 and then inserted it into electrocompetent Escherichia coli. Then we conjugated the electrocompetent cells with wild-type M. aquaeolei to transfer the mutant gene. A ferrozine assay was used to determine M. aquaeolei’s ability to oxidize iron is lost when CCP is mutated; thus our findings show CCP to be a vital component for M. aquaeolei’s ability to oxidize iron. |
| Keyword | iron oxidation; cytochrome c peroxidase; Marinobacter aquaeolei; plasmid construct; mutagenesis |
| Language | English |
| Part of collection | University of Southern California dissertations and theses |
| Publisher (of the original version) | University of Southern California |
| Place of publication (of the original version) | Los Angeles, California |
| Publisher (of the digital version) | University of Southern California. Libraries |
| Provenance | Electronically uploaded by the author |
| Type | texts |
| Legacy record ID | usctheses-m |
| Rights | Waite, Jamie Lynn |
| Access conditions | The author retains rights to his/her dissertation, thesis or other graduate work according to U.S. copyright law. Electronic access is being provided by the USC Libraries in agreement with the author, as the original true and official version of the work, but does not grant the reader permission to use the work if the desired use is covered by copyright. It is the author, as rights holder, who must provide use permission if such use is covered by copyright. The original signature page accompanying the original submission of the work to the USC Libraries is retained by the USC Libraries and a copy of it may be obtained by authorized requesters contacting the repository e-mail address given. |
| Repository name | University of Southern California Digital Library |
| Repository address | USC Digital Library, University of Southern California, University Park Campus MC 7002, 106 University Village, Los Angeles, California 90089-7002, USA |
| Repository email | cisadmin@usc.edu |
| Archival file | uscthesesreloadpub_Volume1/etd-WaiteJamie-602.pdf |
Description
| Title | Page 1 |
| Full text | CHARACTERIZATION OF CYTOCHROME C PEROXIDASE OF MARINOBACTER AQUAEOLEI by Jamie Lynn Waite ________________________________________________________________________ A Thesis Presented to the FACULTY OF THE USC GRADUATE SCHOOL UNIVERSITY OF SOUTHERN CALIFORNIA In Partial Fulfillment of the Requirements for the Degree MASTER OF SCIENCE (BIOCHEMISTRY AND MOLECULAR BIOLOGY) May 2012 Copyright 2012 Jamie Lynn Waite |
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