Bi8980 Protein Preparation and Characterization I - Expression and Purification

Faculty of Science
Spring 2011
Extent and Intensity
2/0. 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)
Guaranteed by
prof. RNDr. Jiří Fajkus, CSc.
Department of Experimental Biology – Biology Section – Faculty of Science
Contact Person: RNDr. Lubomír Janda, Ph.D.
Timetable
Fri 9:00–10:50 B11/306
Prerequisites
C3580 Biochemistry || C3181 Biochemistry I || Bi4010 Essential molecular biology || B4020 Molecular biology || Bi4020 Molecular biology
Basic knowledge of biochemistry and/or molecular biology is a prerequisite for subscription to 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 20 fields of study the course is directly associated with, display
Course objectives
This course is for students to gain the basic knowledge about the preparation of proteins in sufficient quantity and quality for crystallization, NMR analysis and others biochemical measurements. At the end of this course the student will be able to: understand the different approaches in the cloning of various expression vectors using affinity tags or without tags, and subsequent purification of the recombinant protein with affinity chromatography or classically; to remove the fusion protein (anchor); to hold the protein in soluble state; to know various methods for protein detection during the purification procedures.
Syllabus
  • 1. The molecular principles for understanding proteins. 2. Calculation 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 labelling. 12. Invited lecture.
Literature
    recommended literature
  • PRICE, Nicholas C. and Jacqueline NAIRN. Exploring proteins : a student's guide to experimental skills and methods. Oxford: Oxford University Press. xv, 512. ISBN 9780199205707. 2009. info
  • High throughput protein expression and purification : methods and protocols. Edited by Sharon A. Doyle. Totowa, N.J.: Humana Press. xii, 322. ISBN 9781588298799. 2009. info
  • HARDIN, Charles. Cloning, gene expression and protein purification : experimental procedures and process rationale. New York: Oxford University Press. viii, 435. ISBN 0195132947. 2001. info
Teaching methods
The main teaching method are lectures that include particular examples from the own scientific praxis of participating techers and demonstration of the solution of particular problems.
Assessment methods
written test + oral exam
Language of instruction
Czech
Follow-Up Courses
Further Comments
Study Materials
The course is taught annually.
The course is also listed under the following terms Spring 2011 - only for the accreditation, Spring 2010, spring 2012 - acreditation.
  • Enrolment Statistics (recent)
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