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Biosciences Funded PhD - Formation of internal and external lipid vesicles in E.coli for vaccine production

The
studentship will pay an annual stipend at the UK Research Council rate of
£14,553 (rate for 2017/2018) and cover tuition fees at the rate for UK/ EU
students. To be eligible for a full award a student must have no restrictions
on how long they can stay in the UK and have been ordinarily resident in the UK
for 3 years prior to the start of the programme (for EU applicants this
includes time spent studying).

Formation of internal and
external lipid vesicles in E. coli
for vaccine production



Prof
Martin Warren, Prof Mark Smales (Kent), Dr Tarrit Mukhopadhyay (UCL)





We have developed technologies that allow for the formation of both internal
vesicles, derived from the bacterial inner membrane, and external vesicles,
derived from the outer membrane, using E. coli as a chassis organism. In this
project we aim to develop these approaches further to see if the vesicles can
be used for the generation of vaccines.



The
internal vesicles are formed by the expression of the lemA gene, a gene family that also includes mamQ, which is a key component of magnetosome formation. Our
preliminary evidence suggests that the LemA proteins from different organisms
generate different sized internal vesicles and appear to function by pinching
off the membrane. In this respect the cell does not produce a lot of the LemA
protein. We thus now want to determine if we can target proteins to the
internal membranes so that the internal vesicles are rich is a specific protein
of choice. This will be achieved by co-expressing the genes for both LemA and
the specific membrane protein. After growth the cells will be collected, lysed
and the internal membrane vesicles collected by centrifugation. The composition
of the lipid vesicles will then be analysed by a combination of SDS PAGE and
mass spectrometry.



External
vesicles are formed by anchoring the LemA protein to the outer membrane by
fusing it to a BamE lipid anchor. As with the internal membrane system we will
target specific membrane proteins to the outer membranes and then generate the
vesicles in order to determine if the vesicles are enriched with the protein of
choice. This will be done by extracting the vesicles from the cell culture and
analysing them in the same way as described above.

Criteria

The successful candidate is expected to be a highly motivated student.
He/she will be expected to have a minimum of an upper 2nd class degree.

UK residency criteria apply as above.

How to apply

Applications
can be made using the online University application page where the project title should be
entered as the proposed area of research and Prof Martin Warren or Prof Mark
Smales as supervisor. Please include a CV and a cover letter.





Informal
enquiries can be addressed to Prof Martin Warren (m.j.warren@kent.ac.uk) or Prof Mark Smales (c.m.smales@kent.ac.uk)





Deadline

Applications must be received by Friday 14th July 2017


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