C8980 Protein Preparation and Characterization I - Expression and Purification

Faculty of Science
Spring 2023
Extent and Intensity
2/0/0. 2 credit(s) (fasci plus compl plus > 4). Type of Completion: zk (examination).
Teacher(s)
RNDr. Lubomír Janda, Ph.D. (lecturer)
Ing. Blanka Pekárová, Ph.D. (lecturer)
Mgr. Radka Dopitová, Ph.D. (lecturer)
doc. RNDr. Mgr. Jozef Hritz, Ph.D. (lecturer)
Mgr. Adam Norek, Ph.D. (lecturer)
prof. Mgr. Lukáš Žídek, Ph.D. (lecturer)
Guaranteed by
prof. Mgr. Lukáš Žídek, Ph.D.
National Centre for Biomolecular Research - Faculty of Science
Contact Person: RNDr. Lubomír Janda, Ph.D.
Supplier department: National Centre for Biomolecular Research - Faculty of Science
Timetable
Tue 15:00–16:50 C02/211
Prerequisites
Basic knowledge of biochemistry and/or molecular biology is prerequisite for registration for the course.
Course Enrolment Limitations
The course is also offered to the students of the fields other than those the course is directly associated with.
fields of study / plans the course is directly associated with
there are 37 fields of study the course is directly associated with, display
Course objectives
Students completing the course will know the basic theoretical and methodological knowledge to express and purify recombinant proteins. He/she will be able to decide what expression system used for protein expression and what action to take when choosing a purification method. He/she will be able to apply the knowledge to improve the efficiency of expression and the solubility of the proteins. He/she will be able to choose the appropriate method for the detection of proteins, and will be able to prepare label proteins for NMR analysis or crystallography.
Learning outcomes
Graduate student is able to: - analyze protein properties - design a cloning strategy - choose an expression system - select cell disintegration method - design a purification procedure - analyze protein quality - label proteins for structural analysis - express proteins in the bioreactor
Syllabus
  • 1. The molecular principles for understanding the properties of proteins. 2. Calculations in the molecular biosciences 3. The important properties of proteins and how to explore them. 4. DNA cloning. 5. Protein expression. 6. Cell disruption. 7. Fusion proteins and affinity purification. 8. Protein purification chromatography. 9. Protein quality. 10 Analytical methods used to study proteins. 11. Protein labeling. 12. Fermentation technology.
Literature
    recommended literature
  • PRICE, Nicholas C. and Jacqueline NAIRN. Exploring proteins : a student's guide to experimental skills and methods. New York: Oxford University Press, 2009. xv, 512. ISBN 9780199205707. info
  • Protein purification : principles, high resolution methods, and applications. Edited by Jan-Christer Janson. 3rd ed. Hoboken, N.J.: John Wiley & Sons, 2011. xiv, 517. ISBN 9780471746614. info
  • HARDIN, Charles. Cloning, gene expression and protein purification : experimental procedures and process rationale. New York: Oxford University Press, 2001. viii, 435. ISBN 0195132947. info
    not specified
  • Guide to protein purification. Edited by Richard R. Burgess - Murray P. Deutscher. 2nd ed. Boston: Elsevier/Academic Press, 2009. liii, 851. ISBN 9780123749789. info
Teaching methods
The main teaching method are lectures that include particular examples from the own scientific practice of participating techers and demonstration of the solution of particular problems.
Assessment methods
During the semester, students will be tested after each lecture with a written test. In each lesson, students are motivated to get up to 20 extra points, which corresponds to one point on the final test. For the entire semester can earn up to 10 points in the final test (200 extra points). The final test contains thirty test questions with four possible answers. The top ten are have low weight (only one correct answer for one point), the second ten questions have higher weight (maximally two correct answers evaluated with two points) and the last ten questions are worth three points (maximum of three correct answers). Overall, student earn 60 points. Students are graded A if the gains of the test and during the term in the sum of 56 points or more. A grade of F will be given to the student who obtains the sum test and auxiliary points less than 35 points. If the student is not satisfied with the results of a test for him is not an objective criterion of his/her knowledge, the student opt for an oral test with a maximum improvement of the final mark by two grades if demonstrating exceptional knowledge of the issue.
Language of instruction
Czech
Follow-Up Courses
Further Comments
Study Materials
The course is taught annually.
Listed among pre-requisites of other courses
The course is also listed under the following terms Spring 2012, Spring 2013, Spring 2014, Spring 2015, Spring 2016, Spring 2017, spring 2018, Spring 2019, Spring 2020, Spring 2021, Spring 2022.
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