About the Project
The aim of this project is to identify novel bacteriocins produced by clostridia and other bacteria from environmental samples that are likely to harbour bacteria in close association with C. difficile. Biological activity against a range of C. difficile strains including the hypervirulent PCR-ribotype 027 strains will be tested to identify candidate active organisms that can be characterised further to determine the mechanisms of bacteriocin import and activity against sensitive cells.
The work will involve enriching for bacterial colonies from the environmental samples and then replica plating and screening for bacteriocins active against strains of C. difficile by the presence of zones of inhibition around individual colonies on biological activity assay plates. The bacterial colonies will be identified using 16s rDNA sequencing and whole genome sequencing. Activity in cell free extracts will be tested to see if the active bacteriocin is actively secreted into the environment and stable in isolation. We have some preliminary data that suggests expressed bacteriocins have low activity in cell free extracts, but that activity can be significantly enhanced by ammonium sulphate concentration. Concentrated bacteriocins will be investigated further using SDS-PAGE and native PAGE to fractionate samples and allow gel-overlay experiments with sensitive bacterial to identify the active protein. Mass spectrometry of isolated protein bands will be used to identify the candidate protein bacteriocins. In silico analysis of the sequenced genome will allow identification of potential bacterial gene clusters responsible for the expressed active bacteriocin which can be characterised further by gene cloning, protein purification and targeted gene mutations using the well-developed CRISPR-Cas9 system available in the laboratory. Gene engineering may be possible to increase host-range and/or potency of the bacteriocin.
The University of Nottingham is one of the world’s most respected research-intensive universities, ranked 8th in the UK for research power (REF 2014). Students studying in the School of Life Sciences will have the opportunity to thrive in a vibrant, multidisciplinary environment, with expert supervision from leaders in their field, state-of-the-art facilities and strong links with industry. Students are closely monitored in terms of their personal and professional progression throughout their study period and are assigned academic mentors in addition to their supervisory team. The School provides structured training as a fundamental part of postgraduate personal development and our training programme enables students to develop skills across the four domains of the Vitae Researcher Development Framework (RDF). During their studies, students will also have the opportunity to attend and present at conferences around the world. The School puts strong emphasis on the promotion of postgraduate research with a 2-day annual PhD research symposium attended by all students, plus academic staff and invited speakers.
Chikindas, M. L., Weeks, R., Drider, D., Chistyakov, V. A. and Dicks, L. M. (2018). Functions and emerging applications of bacteriocins. Current opinion in biotechnology. 49:23–28. doi: 10.1016/j.copbio.2017.07.011.
Why not add a message here
Based on your current searches we recommend the following search filters.
Based on your current search criteria we thought you might be interested in these.