Lab rotation project description
During the 7 week rotation, the student will express NP and Z proteins of the Lujo strain with a fluorescent tag in mammalian cells. The localisation of these tagged proteins will be analysed by immunofluorescence and the proteins purified from cells in preparation for mass spectrometry. In parallel, NP and Z will be cloned into bacterial expression vectors and expression trials for purification and downstream structural analysis performed.
Week 1: Cell culture techniques and transfection
Week 2: Transfection and cloning
Week 3: Protein expression in mammalian cells,- detection by Immunofluorescence, western blotting
Week 4: Refinement of week 3
Week 5: Purification trials from mammalian cells, coomassie staining, data analysis and write up
Week 6: Expression of proteins in bacteria at different temperatures, coomassie staining
Week 7: Data compilation, analysis and write-up
Training will be provided in cell culture (sterile technique) and protein expression and purification, protein detection methods (immunofluorescence and western blotting) and data analysis. There will also be the opportunity to present the project at lab meetings and to the wider ‘One Virology’ group.