Bi7311 Molecular Biology of Procaryotes - Laboratory Course

Faculty of Science
Autumn 2018
Extent and Intensity
0/2/0. 2 credit(s). Type of Completion: z (credit).
Teacher(s)
Mgr. Ivana Mašlaňová, Ph.D. (seminar tutor)
doc. RNDr. Vladislava Růžičková, CSc. (seminar tutor)
prof. RNDr. Roman Pantůček, Ph.D. (seminar tutor)
Mgr. Adéla Finstrlová, Ph.D. (assistant)
Mgr. Ivana Kupčíková, DiS. (assistant)
Guaranteed by
prof. RNDr. Jiří Doškař, CSc.
Department of Experimental Biology – Biology Section – Faculty of Science
Contact Person: Mgr. Ivana Mašlaňová, Ph.D.
Supplier department: Department of Experimental Biology – Biology Section – Faculty of Science
Timetable
Mon 17. 9. to Fri 14. 12. Tue 10:00–13:50 E25/119, Thu 8:00–9:50 E25/119
Prerequisites
( B2050 General microbiology || Bi2050c General microbiology pr. || B4070 General mikrobiology || B4090 General microbiology || Bi4090c General microbiology - pr. ) && NOW( Bi7120 Molecular biology of prokaryotes )
Basic knowledge from molecular biology of the Prokaryotes. Basic techniques of microbiology and molecular biology.
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 32 student(s).
Current registration and enrolment status: enrolled: 0/32, only registered: 0/32
fields of study / plans the course is directly associated with
Course objectives
The aim of the practice is to acquire knowledge about basic molecular biology methods used for prokaryotic genome analysis, bacterial genetics methods and to understand of principles of horizontal gene transfer. Students will be introduced to identification of changes in prokaryotic genomes due to mutations, horizontal gene transfer via transduction, conjugation and transformation. The practice practically completes the content of lectures from Molecular Biology of Prokaryotes (Bi7120). Students will comprehend the mechanisms of prokaryotic evolution in practice and understand the techniques that can be used to manipulate bacterial genomes.
Learning outcomes
At the end of this course, students should be able to design and perform basic laboratory experiments focused on genetic studies of Escherichia coli and selected species from Gram-positive cocci.
Syllabus
  • 1. Purification of lytic phage particles with therapeutic usage in CsCl gradient.
  • 2. Transduction. The transfer of plasmid resistance genes. Induction of phage phiJB by UV-irradiation, determining the transduction frequency, selection of transductants.
  • 3. Determination of plasmid copy number (PCN) by qPCR.
  • 4. Pulsed-field gel electrophoresis of bacterial genomic DNA (preparation of samples, digestion with restriction endonuclease SmaI, PFGE).
  • 5. Conjugation of Escherichia coli.
  • 6. Electoporation of plasmid DNA to recipient strains. Determining the transformation frequency.
  • 7. Monitoring of phage endolysin expression. Isolation of RNA, transcription of RNA into cDNA, separation of proteins by SDS-PAGE.
  • 8. Isolation of spontaneous mutants of Staphylococcus aureus resistant to antibiotics on gradient agar plates.
Literature
  • SNYDER, Larry and Wendy CHAMPNESS. Molecular genetics of bacteria. 2nd ed. Washington: ASM Press, 2003, xvi, 566. ISBN 155581204X. info
  • MILLER, Jeffrey H. A Short course in bacterial genetics : a laboratory manual and handbook for Escherichia coli and related bacteria. New York: Cold Spring Harbor Laboratory Press, 1992, 456 s. ISBN 0879693495. info
  • ROSYPAL, Stanislav. Obecná bakteriologie. 1. vyd. Praha: Státní pedagogické nakladatelství, 1981, 749 s. info
Teaching methods
The course is taught in research microbiology and molecular biology laboratories. Students perform practical experiments according to the instructions of teacher and according to printed manuals.
Assessment methods
The course is completed after work out an elaborate and pass a writen test.
Language of instruction
Czech
Follow-Up Courses
Further Comments
Study Materials
The course can also be completed outside the examination period.
The course is taught annually.
The course is also listed under the following terms Autumn 2007 - for the purpose of the accreditation, Autumn 2010 - only for the accreditation, Autumn 2002, Autumn 2003, Autumn 2004, Autumn 2005, Autumn 2006, Autumn 2007, Autumn 2008, Autumn 2009, Autumn 2010, Autumn 2011, Autumn 2011 - acreditation, Autumn 2012, Autumn 2013, Autumn 2014, Autumn 2015, Autumn 2016, autumn 2017, Autumn 2019, Autumn 2020, autumn 2021, Autumn 2022, Autumn 2023, Autumn 2024.
  • Enrolment Statistics (Autumn 2018, recent)
  • Permalink: https://is.muni.cz/course/sci/autumn2018/Bi7311