Bacterial Pathogenesis - molecular approach 1400-215MPBP
Course
Terminology (definitions – infection, colonization, disease, virulence factors, opportunistic pathogens, emerging and reemerging pathogens). History of bacteriology. Classical and modern Koch’s postulates. The normal human microbiota (HMP – Human Microbiome Project). Pathogen evolution – HGT (mobile genetic elements), PAIs, evolution by reduction, comparative genomic. Influence of bacterial infections on the development of cancers. Traditional biochemical and genetic approaches and molecular approaches for studying bacterial virulence. Bacterial strategies for evading host defense system (resistance to antibacterial peptides, iron acquisition mechanisms, adherence and invasion, avoiding complements and phagocytosis, evading the host antibody’s response). Delivery of virulence factors – secretion systems. Bacterial adhesion to the cell surface and extracellular matrix of host tissue. Consequences of adhesion – influence on eukaryotic cells signal transductions pathways. Biophilms. Cellular invasion by bacterial pathogens – mechanisms of bacterial entry into host cells, the bacterial life in the cytosol, the bacterial life in the vacuole. Intracellular pathogens ((Shigella, Salmonella, Yersinia sp.). Virulence regulation (phase and antigenic variability, two-component signal transduction system, quorum sensing, small noncoding RNAs). Bacterial exotoxins – structure and function.
Laboratory
During the lab students work in small groups. The lab topics are designed to familiarize students with the basic methods used for genetic and functional analysis of virulence factors and modern methods in diagnostic and genotyping of pathogenic bacterial strains.
The following subjects are included:
1. Identification of genus Campylobacter in environmental samples.
2. Detection of the anty-Helicobacter IgG antibodies in human serum.
3. Analysis of the expression level of genes coding virulence factors (analysis using transcriptional fusions and reporter genes)
4. Functional characteristic of bacterial virulence factors: cellular localization of virulence factors.
5. Analysis of the Dsb genes, responsible for disulphide bond formation.
6. Functional analysis of bacterial virulence factors – construction of the mutant strains - site-directed mutagenesis, insertional mutagenesis.
7. Analysis of selected protein immunogenicity – Western-blot analysis
8. Student familiarization with functioning (procedures, methods, equipments, solved problems) of bacteriological laboratories – field trip to selected laboratories (Department of Molecular Microbiology in The National Medicines Institute (prof. M. Gniadkowski) and/or Department of Clinical Microbiology and Immunology, Department of Microbiology, National Tuberculosis and Lung Diseases Research Institute (prof. Kopeć-Augustynowicz) and/or Department of Genetics, Maria Sklodowska-Curie Memorial Cancer Center and Institute of Oncology (prof. M. Mikula).
Type of course
Mode
Requirements
Prerequisites (description)
Course coordinators
Learning outcomes
Knowledge
Acquires the knowledge of current concepts in major fields of biotechnology, knowledge of the microbiological and genetic terminology (in Polish and English), latest researches, discoveries and utilization of microorganisms in biotechnology, medicine and molecular biology. (K_W02, K_W03 Bt2)
Knows the basic techniques and tools for the study of natural phenomena, especially used to study molecular mechanisms of bacterial pathogenesis. (K_W04 Bt2)
Shows the expertise in designing researches, knows modern data acquisition methods and applications of various laboratory techniques.(K_W05 Bt2)
Shows the knowledge about: (i) independent designing of the scientific experiments, (ii) working out own experimental data for presentations or/and publications, (iii) the significance of experimental work in the field of biotechnology, especially in the field of infectious diseases. (K_W08 Bt2)
Knows and understands the cellular and molecular bases of functioning of prokaryotic microorganisms and the interactions between pathogens and host in respect of research concerning bacterial pathogenesis. (K_W01 Bt1)
Knows how to plan and conduct out genetic modifications of microorganisms using diverse strategies. (K_W04. K_W05 Bt2)
Abilities
Uses the advanced techniques of molecular genetics for selection and targeted modifications of microorganisms.(K_U01 Bt2)
Commands of the modern language (Polish or English) to a degree that enables accessing electronic data sources and research publications concerning bacterial genetic. (K_U02, K_U03 Bt2)
Under the supervision of a scientific tutor is able to design and to perform a simple experiment using the previously learned methods. (K_U04 Bt2)
Is able to explain the results of the experiments and to discuss them on the basis of the accessible literature.(K_U06, K_U07 Bt2)
Social competences
Understands natural biological, physical and chemical phenomena which happen in nature. (K_K01 Bt2)
Appreciates the significance of mathematic and statistic tools used for evaluation of experimental data and phenomena which happen in nature. (K_K02 Bt2)
Is cautious and critical when gaining and interpreting the knowledge concerning bacterial genetics and its application. (K_K04 Bt2)
Understands the need of communication to the society information about the newest achievements in the field of biotechnology, biology of microorganisms and is able to do that in a comprehensive way. (K_K06 Bt1)
Assessment criteria
Assessment methods
The criteria of evaluation of the lab: (i) attendance and active participation (ii) written test. To receive a credit for the lab the student should (i) have no more than two absences during the lab; (ii) receive minimum 51% of the sum of points obtainable in written test (contains 5-6 open questions). The lecture will be assessed on the basis of a multi-choice test (to complete the test one have to receive minimum 51% of the sum of points obtainable in the multi-choice test).
Practical placement
No
Bibliography
Bibliography
Biologia molekularna bakterii. 2006. PWN – chapter 8
Gołąb J. M. Jakóbisiak, W. Lasek. Immunologia. 2002. Wydawnictwo Naukowe PWN
Salyers A.A and D. D. Whitt . 2002. Bacterial pathogenesis. A molecular approach. Second edition. ASM Press. Washington D.C.
Wilson BA. et al.2011. Bacterial pathogenesis. A molecular approach. Third edition. ASM Press. Washington D.C
Sambrook J., Russell D.W. 2001. Molecular cloning - a laboratory manual. 3rd edition. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York – selected chapters
Review articles from scientific journals such as . Nat. Rev. Microbiol.; Curr. Opin. Microbiol.; Trends in Microbiol. FEMS Microbiol. Rev. recomended by academic teacher.
Additional information
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