Docente
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NEGRI RODOLFO
(programma)
Main goal of the course is to give to the students a general view of the molecular mechanisms which are at the base of the biological processes. For each argument a methodological section will introduce the students to the main technical approaches used at the state of the art. Practical training (2 CFU) will make the students to breath the atmosphere of a molecular biology laboratory. Particular attention will be given to the high throughput methods utilized in “Omic” approaches to the comprehension of biological processes which will be further illustrated in the course of Molecular Biology and Genomics.
General aims:
The course is focused on the deep connection between DNA structure and functions. The student will learn the molecular mechanisms which are at the base of the processes of Replication, Recombination, Transcription and DNA repair and their regulatory circuits.
Specific aims:
1. Knowledge and understanding: The student should learn the basic molecular mechanisms of the cellular omeostasis and gene regulation and the most used techniques of Molecular Biology.
2. Ability to apply Knowledge and understanding: The student should be able to apply this knowledge in the discussion of recent arguments of general interest in Molecular Biology research.
3. Critical Judgement abilities: The student should demonstrate critical judgement abilities in solving problems related to theoretical and practical Molecular Biology projects and should communicate its conclusions to the colleagues and to the teacher in an effective way.
4. the student should demonstrate to be able to apply the learned concepts to his future work in molecular biology.
Molecular Biology I
The polimorphic DNA – Chemical components of DNA: bases, sugars and phosphodiesteric backbone (0.1 cfu). DNA B basic structure (0.2). Alternative DNA conformations, unusual structures (crucifiorms, triple helix) (0.2) – Conformational variability of structural parameters, curvature and bendability (0.2). DNA topology, winding and unwinding; linking number; DNA topoisomerases (0.2). The Genetic Code - the code decrypting; the structure and function of the code (0.1). DNA replication: machinery and mechanisms in prokaryotes and eukaryotes (0.2). Replicons organization; topological and end-replication problems (0.1). DNA mutability and repair; damage checkpoints (0.1). DNA transcription: transcription in bacteria and bacteriophages; transcriptional machinery and RNA polymerase positioning signals (0.2). Methodological approach to the study of transcription: in vitro transcription systems (0.2). Transcription regulation in prokaryotic systems: activation and repressions; operon structures and function (0.2). Transcription in eukaryotic systems (0.1); transcriptional machinery and RNA polymerase positioning signals (0.2). Methodological approaches to the study of transcription: eukaryotic in vitro transcription systems. Coordinate regulation of the three eukaryotic RNA polymerases (0.2). Transcription factors and transcription regulation in eukaryotes (0.2). The RNA molecule (chemical and structural features) (0.2); structure and function of tRNA, rRNA, pre- and mRNA, snRNA e snoRNA (0.2); mRNA capping and polyadenilation and their regulative role (0.2). Chromatin basic structure in eukaryotes and prokaryotes (0.2). The structure of nucleosomes and further organization levels (0.2). Histone modifications and their regulatory effects; the histone code. (0.2). DNA methylation and its regulatory role (0.2). Histone variants and their regulatory role. Chromatin structure and chromatin remodelling at promoters (0.2). Chromosomes structure: centromers, telomers and origin of replication (0.1).
Molecular Biology Techniques (0.5 cfu): Nucleic acids purification, quantization, labeling and sequencing (introduction to NGS); PCR, RT-PCR, Southern and Northern blot; basic cloning techniques Techniques for analysis of protein-DNA interactions. Practical training (1 cfu).
James D. Watson, Tania A. Baker, Stephen P. Bell, Alexander Gann, Michael Levine, Richard Losick: Molecular Biology of the Gene
Suggested lectures: selected chapters from: C.R. CALLADINE and H.R. DREW "UNDERSTANDING DNA" - ACADEMIC PRESS
ADDITIONAL MATERIALS (SLIDES AND REVIEWS) WILL BE AVAILABLE ON MOODLE2 PLATFORM
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