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Title: Unraveling the role of small regulatory RNAs (sRNAs) in Clostridium difficile
Author: Joshi, Nimitray
ISNI:       0000 0004 6352 002X
Awarding Body: University of Nottingham
Current Institution: University of Nottingham
Date of Award: 2016
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Clostridium difficile is an opportunistic human pathogen capable of causing a spectrum of diseases ranging from relatively mild antibiotic-associated diarrhoea to severe and life-threatening conditions such as pseudomembranous colitis. Despite its medical importance, comparatively little is known about the regulatory processes that take place in response to environmental changes or to maintain and co-ordinate housekeeping functions. Over the last couple of years, it has become apparent that small RNAs (sRNAs) are playing important regulatory roles in bacteria. Utilizing RNA sequencing (RNA-Seq) technology, the aim of this study was to identify regulatory sRNAs on a Genome-wide scale incorporating both toxin synthesis and sporulation. RNA-Seq data was analyzed using a stream-lined series of Perl scripts and categorized in 1 of 4 groups, according to size, location (within or between genes) and orientation. This yielded a list of thousands’ of candidates, which was narrowed down to 260 possible sRNAs. Several of these were subjected to Northern Blot analysis for further verification. Two confirmed and particularly interesting sRNAs were investigated further, an antisense RNA at the 3' end of grdB and a 6S RNA. The grdB gene encodes a selenoprotein component of the glycine reductase complex. A series of generated mutant strains did not produce the antisense RNA and these need to be analyzed further to fully understand its function. 6S RNA fulfills important roles in many bacteria as a regulator of transcription during stress conditions. However, its inactivation and overexpression in C. difficile resulted in no obvious phenotypic changes Finally, the role of the protein Hfq which interacts with sRNAs was investigated, leading to questions about its essentiality within the organism.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available