C8857c Protein Preparation and Characterization III - practice

Faculty of Science
Spring 2011 - only for the accreditation
Extent and Intensity
0/2/0. 3 credit(s) (plus extra credits for completion). Type of Completion: graded credit.
Teacher(s)
doc. Mgr. Lumír Krejčí, Ph.D. (lecturer)
Guaranteed by
doc. Mgr. Lumír Krejčí, Ph.D.
National Centre for Biomolecular Research – Faculty of Science
Prerequisites
It is highly recommended to participate in other courses that are part of Protein Preparation and Characterization.
Course Enrolment Limitations
The course is also offered to the students of the fields other than those the course is directly associated with.
The capacity limit for the course is 10 student(s).
Current registration and enrolment status: enrolled: 0/10, only registered: 0/10, only registered with preference (fields directly associated with the programme): 0/10
fields of study / plans the course is directly associated with
Course objectives
At the end of the course students should be able to understand basic methodology to monitor or characterize protein-mediated interactions. The spectrum of methods will include study of protein-protein interaction by in vitro pull-down assay using purified proteins or in vivo using yeast 2-hybrid method. Example of complex protein interactions will be presented on posttranslational modification by SUMO protein. Protein-DNA interactions will be monitored by electrophoretic mobility shift assay. To study mobile enzymes on a DNA, the ability of helicases to translocate on separate DNA strands will be monitored.
Syllabus
  • 1. Protein-protein interactions and post-translational modification a) pull-down b) sumoylation c) two-hybrid 2. Protein-DNA interaction and 2-hybrid a) EMSA b) 2-hybrid c) data evaluation 3. Protein-DNA interaction a) helicase assay b) data evluation
Literature
  • MEYERS., Robert A. Encyclopedia of Molecular Cell Biology and Molecular Medicine, 2nd Edition. 2004. ISBN 3-527-30545-9. info
Teaching methods
Each day will start with theoretical section describing the background and use of the individual methods. The students will be then divided in group of two and they will perform all tasks practically based on available protocol.
Assessment methods
Each practical course will be evaluated based on the written protocols including control question. To pass the practical course each students has to successfully complete 3/4 of all experiments.
Language of instruction
English
Follow-Up Courses
Further comments (probably available only in Czech)
The course is taught annually.
The course is taught: in blocks.
The course is also listed under the following terms Spring 2010, Spring 2011, Spring 2012, spring 2012 - acreditation, Spring 2013, Spring 2014, Spring 2015, Spring 2016, Spring 2020, Spring 2021, Spring 2022, Spring 2023, Spring 2024.