The course will provide hands-on experience of high throughput (HTP) protein expression screening with an emphasis on methods to study "difficult" targets, including complexes and membrane proteins. Construct design, HTP cloning and protein expression screening in both E. coli and eukaryotic cells will be covered through lectures and practicals as follows :-
1) Designing expression constructs (lecture & in silico workshop).
2) Transient transfection of HEK cells (lecture & practical).
3) Construction of baculoviruses and testing expression (lecture & practical).
4) Screening membrane protein expression using in-gel fluorescence and fluorescence detected size exclusion chromatography (FSEC) (lecture & practical).
A tour of the Diamond Synchrotron and visit to the Membrane Protein Laboratory will also be arranged.
1) Designing expression constructs (lecture & in silico workshop).
2) Transient transfection of HEK cells (lecture & practical).
3) Construction of baculoviruses and testing expression (lecture & practical).
4) Screening membrane protein expression using in-gel fluorescence and fluorescence detected size exclusion chromatography (FSEC) (lecture & practical).
A tour of the Diamond Synchrotron and visit to the Membrane Protein Laboratory will also be arranged.
