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Spatiotemporal pathogen-host interactions during African swine fever virus infection

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  • Full or part time
    Dr C Netherton
    Dr C Benfield
  • Application Deadline
    Friday, March 29, 2019

Project Description

African swine fever virus causes a severe disease of domestic pigs that is spreading at an alarming rate throughout Europe and China. Little is known about the interactions between ASFV and host proteins during infection including that of the cellular receptor used for ASFV entry. The Pirbright Institute has set up stimulated emission depletion (STED) super resolution microscopy as a tool to study African swine fever virus replication and propose to combine this with the Friedrich-Loeffler-Institut’s expertise in proteomics to gain a detailed understanding of the virus-host interactions of key viral proteins.

The successful applicant will generate recombinant viruses encoding proteins fused to tags suitable for spatiotemporal analysis by super resolution microscopy (Eckhardt, 2011) and affinity purification and mass-spectrometry (AP-MS; Gerold, 2016). These viruses will then be used to study the localisation of tagged viral proteins in live cells over time using super resolution microscopy and identify key events such as the formation of the viral replication sites or entry into the host cell. Viral and cellular host interacting partners at different time points will then be identified by AP-MS at Friedrich-Loeffler-Institut and further analysed at Pirbright.

This approach will lead to a greater understanding of the cellular factors involved in formation of African swine fever virus replication complexes and entry into the host cell. It will also represent a proof-of-principle to study the interactions of any of the 150 ORFs encoded by African swine fever virus. The successful applicant will work in the African swine fever vaccinology group at Pirbright and be registered with Camilla Benfield at the Royal Veterinary College. The lab work will take place in the high containment laboratories at The Pirbright Institute with a six to twelve month placement at the Friedrich-Loeffler-Institut near Griefswald, Germany.

Funding Notes

This is a fully funded studentship open to science graduates (with or who anticipate obtaining at least 2.1 or equivalent in relevant biological subject in undergraduate degree, or a Masters degree - subject to university regulations). Open to UK students and eligible EU students who qualify for home-rated fees in line with Residential Guidelines for Research Council studentships – see Apply Online for details. Eligible students will receive a minimum annual stipend of £15,009; university registration fees will be paid. Students without English as first language must provide evidence of IELTS score of 7.0, no less than 6.5 in subsections.

References

1. Eckhardt M, Anders M, Muranyi W, Heilemann M, Krijnse-Locker J, Müller B. (2011) A SNAP-tagged derivative of HIV-1--a versatile tool to study virus-cell interactions. PLoS One. 6(7):e22007. doi: 10.1371/journal.pone.0022007
2. Gerold G, Bruening J, Pietschmann T. (2016) Decoding protein networks during virus entry by quantitative proteomics. Virus Res. 218:25-39. doi: 10.1016/j.virusres.2015.09.006.
3. Keßler C, Forth JH, Keil GM, Mettenleiter TC, Blome S, Karger A (2018) The intracellular proteome of African swine fever virus. Sci Rep. 8(1):14714. doi: 10.1038/s41598-018-32985-z.



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