During the block course in the fall semester, we will carry out biological-chemical enzyme evolution experiments using molecular genetic mutation technologies and in vivo selection in recombinant bacterial strains. The class with its very dense program consists of the practical course itself and an integrated series of seminar/lecture sessions.
Learning objective
All technologies used for the experiments will be explained to the students in theory and in practice with the goal that they will be able to independently apply them for the course project and in future research endeavors. After the course, an individual report about the results obtained has to be prepared.
Content
The class deals with a specifically designed and genuine research project. We intend to carry out biological-chemical enzyme evolution experiments using molecular genetic mutation technologies and in vivo selection in recombinant bacterial strains. By working in parallel, teams of 2 participants each will generate a variety of different variants of a chorismate mutase. Individual enzyme catalysts will be purified and subsequently characterized using several different spectroscopic methods. The detailed chemical-physical analyses include determination of the enzymes' kinetic parameters, their molecular mass, and the integrity of the protein structure. The results obtained from the individual evolution experiments will be compared and discussed at the end of the class in a final seminar. We expect that during this lab course we will not only generate novel enzymes, but also gain new mechanistic insights into the investigated catalyst.
Lecture notes
A script will be distributed to the participants on the first day of the course.
Literature
General literature to "Directed Evolution" and chorismate mutases, e.g.:
– Taylor, S. V., P. Kast & D. Hilvert. 2001. Investigating and engineering enzymes by genetic selection. Angew. Chem. Int. Ed. 40: 3310-3335.
– Jäckel, C., P. Kast & D. Hilvert. 2008. Protein design by directed evolution. Annu. Rev. Biophys. 37: 153-173.
– Roderer, K. & P. Kast. 2009. Evolutionary cycles for pericyclic reactions – Or why we keep mutating mutases. Chimia 63: 313-317.
Further literature will be indicated in the distributed script.
Prerequisites / Notice
This laboratory course will involve experiments that require a tight schedule and, particularly in the second half, very long (!) working days. The maximum number of participants for the laboratory class is limited, but surplus applicants may contact P. Kast directly to have their names added to a waiting list. A valid registration is considered a commitment for attendance of the entire course, as involved material orders and experimental preparations are necessary and, once the class has started, the flow of the experiments must not be interrupted by individual absences. In case of an emergency, please immediately notify P. Kast. For more information see http://www.kast.ethz.ch/teaching.html, from where you can also download a flyer.
Performance assessment
Performance assessment information (valid until the course unit is held again)
Repetition only possible after re-enrolling for the course unit.
Additional information on mode of examination
Credit points can only be earned if the candidate (i) actively takes part in the entire program, (ii) turns in a final report, and (iii) obtains at least grade 4 (4 = satisfactory; scale 1-6; 6 being the best grade). The grade reflects the performance of each participant and is based on (1) effort/participation/presentation (30%), (2) skills or acquisition of skills during the course (30%), and (3) quality of the report (40%). Guidelines for writing the report are available in the section "Learning materials".
Learning materials
No public learning materials available.
Only public learning materials are listed.
Groups
No information on groups available.
Restrictions
General
Permission from lecturers required for all students
Performance assessment as a semester course