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Dissertation
Identification and characterization of novel mutants of Campylobacter jejuni
Doctor of Philosophy (Ph.D.), Drexel University
Jun 2012
DOI:
https://doi.org/10.17918/etd-3787
Abstract
Campylobacter jejuni is one of the most prevalent diarrheal diseases worldwide, having a major effect on human health both in developed and developing countries. While it can be difficult to grow and manipulate in the lab, it remains persistent in our food supply. Mechanisms of virulence in humans have begun to be elucidated with the help of completed genome sequences and development of molecular research tools. Despite this, knowledge of C. jejuni genetics and virulence still trails behind many other intracellular pathogens. In the work presented here a signature tagged mutagenesis system previously created for C. jejuni was implemented and a quantitative real-time PCR based system for detection of the mutagenesis system was developed. The mutagenesis system was then used to identify two novel characteristics of C. jejuni. First, a novel red growth phenotype was identified in C. jejuni which responded to adenine supplementation the same as red yeast mutants and was always associated with a decrease in virulence. Second, a major restriction barrier in C. jejuni strain NCTC11168 was identified. We identified one gene, cj1051c, which when knocked out allowed a 1000-fold increase in transformation efficiency with plasmid purified from a C. jejuni host, as well as permitted transformation with plasmid purified from an E. coli host. In this work we implemented an existing signature tagged mutagenesis system and constructed a novel detection system allowing the quantification of many random mutants in a single screen. This system was then used to identify two novel characteristics of C. jejuni providing an increased knowledge of C. jejuni physiology and virulence as well as potential tools for future research.
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Details
- Title
- Identification and characterization of novel mutants of Campylobacter jejuni
- Creators
- Jeffrey Paul Holt - DU
- Contributors
- Jennifer J. Quinlan (Advisor) - Drexel University (1970-)
- Awarding Institution
- Drexel University
- Degree Awarded
- Doctor of Philosophy (Ph.D.)
- Publisher
- Drexel University; Philadelphia, Pennsylvania
- Resource Type
- Dissertation
- Language
- English
- Academic Unit
- Biology; College of Arts and Sciences; Drexel University
- Other Identifier
- 3787; 991014632573204721