The Construction and Use of a Francisella tularensis DNA Microarray

LeButt, Helen (2008). The Construction and Use of a Francisella tularensis DNA Microarray. PhD thesis The Open University.



A DNA microarray was designed and constructed using the genome sequence of the highly virulent obligate intracellular pathogen Francisella tularensis strain Schu S4. The microarray was optimised and then tested by performing a comparative genomics study on Francisella strains. The microarray was used to distinguish between Francisella strains at the subspecies level, detecting differences between the genomes of the subspecies at a similar rate to differences previously published from Francisella comparative genomics studies. Further analysis of the genomic differences identified between subspecies using the microarray has provided some suggestions as to the genetic basis for the relative attenuation of one subspecies, and similarly, differences identified between the F. tularensis live vaccine strain and its progenitor strain provided some clues as the genetic basis for the attenuation of the vaccine strain. The microarray was also used to carry out functional genomics studies on Francisella novicida cultured under in vitro stress conditions: iron starvation, oxidative stress, elevated temperature, and acidic pH. A number of genes were regulated in response to each of these conditions, and a detailed analysis of the data has provided insights into the stress response of Francisella, and some of the mechanisms that it may employ upon encountering similar stresses in vivo.

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