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529-0044-00L 6 Credits BSC D-CHAB , D-BIOL
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Genetic Code Expansion for Studying Posttranslational Modifications

Lecturers & Examiners: Prof. Dr. Kathrin Lang, Dr. Maximilian Fottner
Enrolment limited to max 12 students. The enrolment is done by the D-BIOL study administration.
VVZ CR n/a

Last Updated: 2026-06-01 11:30:38

Abstract

In this block course we will prepare proteins bearing site-specifically installed post-translational modifications using genetic code expansion. The impact of the post-translational modifications on enzymatic activity and protein-protein interactions will be studied.

Objective

Students will learn basic techniques for genetic code expansion. They will learn how to synthesize non-canonical amino acids (ncAAs), manipulate plasmids through cloning techniques to introduce an amber codon at a user defined site in a gene of interest, express and purify recombinant proteins bearing site-specific ncAAs using state-of-the-art chromatographic techniques, characterize the target protein using mass spectrometry and analyze the properties of the target protein regarding enzymatic activity and protein-protein interactions using biophysical techniques. After the course, participants should be able to apply the gained skills in future chemical and molecular biology lab courses and projects. Individual reports describing the experiments and obtained results must be prepared by the end of the course. At the end of the course, each team (consisting of two participants each) will present their results in an oral presentation.

Content

During this block course, the participants will study the impact of different post-translational modifications (PTMs) on protein function using genetic code expansion. The participants will work in teams of two, will first synthesize different ncAAs mimicking PTMs, such as acylated lysine derivates (e.g. acetlylysine, butryllysine etc.). These ncAAs will be incorporated into target proteins using orthogonal aminoacyl tRNA synthetase / tRNA pairs, which direct the site-specific incorporation in response to an in frame introduced amber stop codon (amber suppression). Therefore, the participants will introduce the amber stop codon at different positions in the gene of interest using standard molecular cloning techniques. After recombinant expression of the target protein, the participants will isolate the ncAA-bearing target proteins using different chromatography techniques using a FPLC system. After characterization of the PTM-bearing protein using mass spectrometry, the participants will study the impact of the PTM on enzymatic activity and protein-protein interactions using biophysical techniques.

Resources

Lecture Notes

All the required theoretical and experimental informations can be found in the provided script. Details will be discussed in class. Relevant research papers will be recommended for reading during the seminars.

General Information

Language
English
Levels
BSC
Frequency
Yearly recurring

Examination

Type
graded semester performance
Student performance will be evaluated by a written report and an oral presentation. Students are obliged to be present throughout the block course.Credit points can only be earned if the candidate (i) actively takes part in the entire 3.5-week program, (ii) turns in a final report, and (iii) obtains at least grade 4.0 (scale 1-6; 4 = pass; 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 will be communicated at the beginning of the block course.Cancellation: If you have to deregister from a course that has been assigned to you (just emergency reasons), please notify in written the course coordinator at least four weeks before the course starts, for courses in the 1st quarter, a cancellation period of one week applies. The study secretariat D-BIOL must also be informed (email CC [email protected]) so that the enrollment is deleted. Otherwise the course is considered as "failed".

Course Components

Type Title Time & Place Hours
practical/laboratory course Genetic Code Expansion for Studying Posttranslational Modifications
Block course in the 2nd quarter of the autumn semester.
No time listed 100 h semesterly

Offered In