Bi8312 Practical Course in Molecular Biology of Viruses

Faculty of Science
Spring 2025
Extent and Intensity
0/2/0. 2 credit(s). Type of Completion: z (credit).
Teacher(s)
Mgr. Tibor Botka, Ph.D. (seminar tutor)
Mgr. Ing. Eliška Kučerová (assistant)
Guaranteed by
Mgr. Tibor Botka, Ph.D.
Department of Experimental Biology – Biology Section – Faculty of Science
Contact Person: Mgr. Tibor Botka, Ph.D.
Supplier department: Department of Experimental Biology – Biology Section – Faculty of Science
Prerequisites
Bi6400 Methods of molecular biology
1. Basic methods in molecular biology; 2. Molecular biology of DNA and RNA viruses; 3. Life and reproduction of bacteriophages.
Course Enrolment Limitations
The course is only offered to the students of the study fields the course is directly associated with.

The capacity limit for the course is 15 student(s).
Current registration and enrolment status: enrolled: 0/15, only registered: 8/15
fields of study / plans the course is directly associated with
Course objectives
The course includes preparation and cultivation techniques of bacterial viruses: Preparation of phage lysate; Determination of the virion titer; One-step growth curve and phage burst size determination by quantitative PCR; Determination of mutation index; Induction of prophages using mitomycin C; Isolation of spontaneous kayvirus mutants on resistant Staphylococcus aureus strains; comparative analysis of changes in viral genomes by PCR, sequencing, and bioinformatics; Purification and concentration of viral particles. After graduation this course, students should be able to conduct and fully understand experiments, from the acquisition of input material to the analysis and interpretation of experimental data.
Learning outcomes
At the end of the course students should be able to:
• isolate viral particles and DNA in sufficient quality and quantity
• analyze isolated genetic material by methods of molecular biology such as PCR, qPCR, ELFO, DNA sequencing, and bioinformatics tools
• isolate and prepare lysate of spontaneous viral mutants effective on up-to-now phage-resistant bacterial strain
• determine and compare the growth properties of bacterial viruses
• determine and compare the growth properties of lysogenic strains depending on the type of prophage and the mode of its induction used
• use theoretical knowledge of other methods and principles in subsequent laboratory practice
Syllabus
  • 1. Isolation of spontaneous mutants of bacterial viruses, determination of mutation frequency and sequence analysis 2. Determination of susceptibility of Staphylococcus aureus strains to polyvalent Kayvirus mutants 3. One-step growth curve and adsorption curve of staphylococcal virus 812 and its mutants, determination of burst size by qPCR 4. Observation of growth properties of lysogenic bacteria using spectrophotometer, induction using mitomycin C and isolation of resident bacterial viruses 5. Purification of viral particles by various ultrafiltration techniques
Literature
  • Návody k cvičení jsou u vyučující
  • Virology methods manual. Edited by B. W. J. Mahy - Hillar O. Kangro. London: Academic Press, 1996, x, 374. ISBN 0124653308. info
  • SAMBROOK, J., E.F. FRITSCH and T. MANIATIS. Molecular Cloning. A laboratory Manual. Second Edition. Cold Spring Harbor: Cold Spring Harbor Laboratory Press, 1989. ISBN 0-87969-309-6. info
  • ROSYPAL, Stanislav. Obecná bakteriologie. 1. vyd. Praha: Státní pedagogické nakladatelství, 1981, 749 s. info
Teaching methods
This course runs as a practical exercise. Students conduct the experiments in assignments ranked according to course syllabus. Study materials are available in the IS.
Assessment methods
The course is successfully completed if the student completes all exercises, passes a written test and submits all protocols.
Language of instruction
Czech
Further comments (probably available only in Czech)
The course is taught annually.
The course is taught: in blocks.
Teacher's information
Students would independently manage the entered tasks and corectly interpret the results in laboratory protocols.
The course is also listed under the following terms Spring 2008 - for the purpose of the accreditation, Spring 2011 - only for the accreditation, Spring 2003, Spring 2004, Spring 2005, Spring 2006, Spring 2007, Spring 2008, Spring 2009, Spring 2010, Spring 2011, Spring 2012, spring 2012 - acreditation, Spring 2013, Spring 2014, Spring 2015, Spring 2016, Spring 2017, spring 2018, Spring 2019, Spring 2020, Spring 2021, Spring 2022, Spring 2023, Spring 2024.
  • Enrolment Statistics (recent)
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