MOLECULAR MICROBIOLOGY
Academic year and teacher
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- Versione italiana
- Academic year
- 2021/2022
- Teacher
- SILVIA SABBIONI
- Credits
- 6
- Curriculum
- Biologia molecolare e cellulare
- Didactic period
- Secondo Semestre
- SSD
- BIO/19
Training objectives
- To provide the basis of microbial genetics in order to understand their industrial and biotechnological use. Microrganisms are studied as models for understanding biological processes. To provide knowledge to understand the pathogenic, diagnostic and preventive aspects related to SARS-CoV-2. To provide the knowledge to understand the mechanisms of virus-host interaction in human oncogenesis. To provide the knowledge for the use of viral vectors in experimental models in vitro and in vivo.
Knowledge and understanding
The student:
- knows molecular, pathogenic, diagnostic and prevention aspects related to SARS-CoV-2;
- knows aspects related to the industrial use of microorganisms;
- knows the molecular basis of cell transformation and tumorigenesis associated to viruses;
- knows the viral vectors and their use.
- Can describe the interaction between microRNA and cellular and viral mRNAs;
- Can describe different regulation mechanisms of viral vectors replication
Applying knowledge (ability)
The student:
- knows how to correctly use scientific information relating to the SARS-CoV-2 pandemic, and is potentially able to contribute to their correct dissemination;
- knows how to evaluate the possible impact of guest- microorganism interaction on the variation of cellular functions during the process of transformation;
- is able to use that knowledge to program a laboratory experiment;
- is able to use of the main techniques for the identification of possible targets of viral microRNA. Prerequisites
- The student must have a knowledge of the basics of microbiology.
Course programme
- The course consists of lectures and laboratory activities.
The lectures will cover the following topics:
Respiratory Viruses, Coronaviruses and SARS-CoV-2: (8 hours)
General information on respiratory viruses and coronaviruses. SARS-CoV-2: origin, epidemiology, structure, replication, pathogenicity. Diagnostic methods (antigenic tests, molecular tests based on the identification of nucleic acids, rapid antigenic tests). Prevention methods, including vaccines. General information on vaccines and in particular vaccines for the prevention of severe COVID-19. Onset of variants and possible consequences.
Bacterial genetics and transmission of acquired property through horizontal gene transfer (TGO): (10 hours)
Acquisition of new properties through mutations; auxotrophic mutants; screening and selection; reversion; application tests based on reversion: the Ames test; industrial use of mutants: selection of a lysine-producer strain.Transformation: the concept of competence, characteristics.Generalized and specialized transduction. Plasmids and Conjugation. Industrial production of new generation vaccines (whole cell or subunit, recombinant). Development of antibiotic resistance: molecular mechanisms of resistance to several classes of antibiotics.
Microorganisms and oncogenesis: examples (12 hours)
molecular mechanisms and virulence factors involved in the infectious process and in gastric cancer associated with Helicobacter pylori. Molecular mechanisms of virus replication, in particular papillomavirus, polyomavirus and the hepatitis viruses. Recombinant vaccines anti HBV and HPV. Pathogenetic mechanisms, especially virus-related oncogenesis.
Microorganisms and oncogenesis: viral vectors in experimental models in vitro and in vivo. (10 hours)
Development and use of plasmid and viral vectors. Study of the functions of viRNA and miRNA: methods to identify target sequences; development and use of viral vectors for the expression of miRNAs and target sequences in vitro and in vivo. Development of viral vectors for new anti-cancer therapeutic approaches.
Laboratory activities: (12 hours)
• Transformation of competent E.Coli with a recombinant DNA, selection of transformed E.Coli cells on ampicillin- containing agar plates; purification of plasmid DNA, and analysis by restriction enzyme digestion.
• Transfection of the recombinant DNA in eukaryotic cells; evaluation of microRNA-target interaction in the transfected cell: analysis of the expression of the transgene by luciferase test using a luminometer. Interpretation of results. Didactic methods
- The course consists of theory lectures and laboratory activities in the microbiology laboratory. In particular, it provides a total of 52 hours (5+1 credits) including 40 hours of lectures and 12 hours of laboratory activities. Classes are held weekly in the classroom, using power-point slides.
For laboratory activities the students are divided into groups (maximum 30 students per group). The students will be guided by the teacher in all practical activities that will be proposed, and will be accompanied to the understanding and correct interpretation of the results obtained from the laboratory activity. Learning assessment procedures
- The aim of the learning assessment procedures is to verify the level of knowledge and deepening of the topics of the course program and the reasoning skills developed by the student. The assessment is expressed in thirtieths (minimum grade 18). With the exception of students who have difficulties, the exam consists of written answers to 20 open questions that require short and precise answers, in order to check the level of understanding and knowledge of the topics of the program. The answers to the questions are judged for both the content and the scientific language, and the maximum score for answer is 5 points. In case of a rating equal to or less than 2 in two or more answers, the examination is not passed. Evaluation of the laboratory activities will focus on the ability to present and to interpret the experimental results.
Reference texts
- Diagrams and figures provided by the teacher.
For consultation and referring to viral vectors for gene therapy:
Giacca Mauro Terapia Genica ed Springer
For consultation and with respect to other aspects of molecular microbiology:
Dehò, Galli Biologia dei Microorganismi, ed Zanichelli
