MRes - Towards efficient selection of attenuated bacterial vaccine strains against enzootic pneumonia
Supervisors: Dr Rob Noad and Professor Dirk Werling
Department: Pathobiology and Population Sciences
Project Details
Enzootic pneumonia is one of the most common respiratory diseases in pigs. Current vaccines are not able to stop the circulation of the causative bacterial pathogen (Mycoplasma hyopneumoniae) within the population. Advances in the genetic manipulation of M.hyop made at RVC have allowed the generation of attenuated bacteria by transposon mutagenesis. However, these mutants all carry an antimicrobial resistance gene and this represents a limitation on their use as live vaccines in livestock. An ideal mutant would be attenuated in pigs through gene interruption or deletion but would not carry any additional antimicrobial resistance. To address this problem we have generated transposon variants that allow the removal of the antibiotic resistance gene by directed recombination. However the remaining technical problem is that it is necessary to select antibiotic sensitive bacteria on a background of antibiotic resistant bacteria. This is quite inefficient, M.hyop colonies are slow growing on plates and the antibiotic selection we are using at the moment works far better in liquid culture than on the plate format. A far better system would be to incorporate a counterselectable marker with the transposon to allow active selection of mutants which had lost the antimicrobial resistance gene.
This MRes project is aimed at assessing the feasibility of using counterselectable markers as a way to select attenuated M.hyop where antibiotic resistance genes have been removed. By the end of the project the student will have completed a literature review and tested a subset of selectable markers for use in M.hyop. We have already identified some genes that could potentially be used for counterselection or to select for the loss of the antimicrobial resistance gene but part of the MRes would be for the student to analyse the literature and to prioritize which approaches are tested first. This is aligns with a larger ongoing BBSRC funded project at the college which is aimed at developing attenuated vaccines to the pathogen.
A student undertaking the research would be part of an active laboratory based mycoplasma research team and would learn skills relating to: experimental design, molecular microbiology, design and selection of synthetic gene sequences, DNA sequence analysis, working with genetically modified bacteria, data analysis and presentation.
References
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Reyrat et al., 1998. https://doi.org/10.1128/iai.66.9.4011-4017.1998
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Maglennon et al., 2013a. https://doi.org/10.1186/1297-9716-44-124
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Maglennon et al 2013b. https://doi.org/10.1186/1297-9716-44-63
Requirements
Essential:
- Must meet our standard MRes entry requirements.
- Must have an undergraduate class 2:I degree in biological sciences or related subject (veterinary degree would be acceptable).
Desirable:
- 3 months laboratory experience with bacteria or molecular techniques.
- Experience of working with at least one counterselectable marker.
- Can demonstrate ability to work in a team.
- Can demonstrate the ability to undertake a research project.
- Can critically analyse the research literature.
This can be taken full-time or part-time (12months FTE) project commencing in October 2024, based at RVC's Hawkshead campus.
Funding
Partially funded: The lab will be covering the project costs, with the MRes student expected to meet the course fees and their living expenses.
International applicants are welcome to apply but must be able to fund the difference between "Home" and "Overseas" tuition fees. Please note that EU/EEA and Swiss national students may no longer be eligible for the “Home” rate of tuition fees, dependent on personal circumstances (including immigration status and residence history in the UK) and UK government rules which are currently being developed. For up-to-date information on fees for EU/EEA and Swiss national students following Brexit please see our fees and funding page.
How to Apply
For more information on the application process and English Language requirements see How to Apply.
Deadline: Sunday 14th July 2024
We welcome informal enquiries - these should be directed to Dr Rob Noad (rnoad@rvc.ac.uk)
Interview date and location: TBC