3 Objectives for Week 1
- Laptops will be used for reading and understanding the background information on expressing proteins containing non-canonical amino acids (ncAAs).
- Obtain and discuss with peers information on the genes and sites of ncAA incorporation available this term.
- Determine the amino acid sequence of the proteins and ncAA locations.
- Register to use and install the free educational version of Pymol at http://pymol.org/edu/
- Download a crystal structure of the proteins from the online Protein Data Bank.
- Load and study the protein structure in Pymol (for help getting started see http://www.pymolwiki.org/index.php/Practical_Pymol_for_Beginners as well as the PyMol Tutorial posted on Canvas).
- Consider which location and which ncAAs will be incorporated/studied based on literature searches and the Pymol image. Identify sites that have already been mutated. Finalize selection of which protein you will study this term and begin to develop hypothesis for term-long projects.
- Set up group bench and calibrate and sterilize pipettes.
- Obtain instructions on expressing protein from the plasmid-containing cells.
- Ensure that the necessary media component solutions and sterile equipment are prepared and available for expressions during week 2 (wild-type protein) and week 3 (set of wild-type and ncAA-mutant proteins).