PhD-Course
26.03-27.03.2015
(max. 6 participants)
Saccharomyces cerevisiae - a versatile tool to study transport proteins
The aim of this course is to get experimental experience with the heterologous
expression system yeast. The yeast expression system can be used for various
transport assays characterizing plant transport proteins.
Day 1:
Morning:
- Lecture1: Theory of yeast complementation and toxicity growth assays
- Preparations of buffers, media etc.
- Yeast uptake assay for elemental analysis (boric acid)
Afternoon:
- Yeast complementation and toxicity growth assays (boric acid, urea,
ammonia, hydrogen peroxide ...)
Day 2:
Morning:
- Interpretations of the yeast complementation / toxicity growth assays
and the elemental uptake study (prepared assays analog to the performed
ones)
- Lecture2: Theory of stopped flow measurements
- Yeast protoplast preparation
Afternoon:
- stopped-flow water uptake assays (protoplast swelling)
- stopped-flow ammonia or hydrogen peroxide uptake assays (fluorescent
dye)
.... = theoretical work
.... = practical work