1. Knowledge and understanding. Students will need to know the physiological structures of nucleic acids and the functioning of prokaryotes and eukaryotes with the molecular basis of the flow of genetic information. Students will also need to know the applications of molecular biology to biotechnology. 2. Ability to apply knowledge and understanding. At the end of the course the students will have to know and be able to correlate the fundamental processes of molecular biology to the cellular functions in prokaryotes and eukaryotes. Students will need to be able to apply the principles of molecular biology to biotechnologies and to the techniques of studying genes and gene expression. 3. Autonomy of judgment. At the end of the course the students will be able to connect the mechanisms of DNA modulation (synthesis, repair, transcription and translation) for the regulation of gene expression in response to environmental stimuli in prokaryotic and eukaryotic cells. Moreover, students will have to be able to explain the molecular principles on which the technologies are based. 4. Communication skills. At the end of the course the students will have to be able to expose the fundamental processes of the molecular biology of the living and its applications with appropriate scientific terminology and with clarity. 5. Learning ability. Students will have to demonstrate that they have learned from listening to the lessons, from the teaching material provided and from discussions with the teacher. Students will have to demonstrate an independent and critical ability to interpret biological phenomena with the principles of molecular biology.

Basic knowledge of biochemistry

The cell and the sizes of organisms. The difference between prokaryotes and eukaryotes. The organelles of eukaryotic cells. Cell duplication. The discovery of DNA as fundamental genetic material. The discovery of the DNA double helix. The secondary structures of DNA. Base tautomerism, keto-enol and amino-imine shifts and the possibility of fixing mutations. DNA stability and base hyperchromism: Tm. DNA renaturation kinetics. The complexity of genomes and the value of Cot. The RNA world. The best-known local structural motifs. The ribozymes. DNA and its different topological forms. Topoisomerases. Chromatin and the compaction of DNA into fibers. The role of histone tails in chromatin remodeling. The size of the genomes of prokaryotic and eukaryotic organisms. The three paradoxes of genomes: K, N and C. Human DNA. The sizes of introns and exons in different organisms. MiRNAs: biogenesis and their role. piRNAs and circRNAs. Gene families. The chemistry of replication. The structure of bacterial DNA polymerase. The replication fork and the synthesis of the continuous and discontinuous filament. The phases of replication: initiation, elongation and termination. ORIs and consensus sequences. The DNA pol of eukaryotes. The RFC and the PCNA. RPA and DNA primase. The removal of primers in eukaryotes. Biological significance of telomeres. Nucleosomal inheritance (outline). DNA damage and mutations. Chromosome aberrations. Genetic diseases and somatic diseases. DNA repair systems in prokaryotes and eukaryotes: similarities and differences. DNA transcription in prokaryotes and eukaryotes: the types of transcribed RNA and RNA polymerase. The mono and polycistronic gene. The stages of transcription: beginning, progress and end. The splicing process and intron removal. The genetic code and its universality. Translation and its phases: comparison between prokaryotes and eukaryotes. Protein folding and their compartmentalization. Recombinant DNA technology: gene cloning and protein expression. Molecular techniques for the study of genes and gene expression.

Zanichelli: Bruce Alberts, Rebecca Heald, Alexander Johnson, David Morgan, Martin Raff, Keith Roberts, Peter Walter Biologia molecolare della cellula, 7e 2025, pagine 1520 Lizabeth A. Allison Fondamenti di biologia molecolare, 2e 2023, pagine 592 Harvey Lodish, Arnold Berk, Chris A. Kaiser, Monty Krieger, Anthony Bretscher, Hidde Ploegh, Kelsey C. Martin, Michael B. Yaffe, Angelika Amon Biologia molecolare della cellula, 4e 2022, pagine 1200 James D. Watson, Tania A. Baker, Stephen P. Bell, Alexander Gann, Michael Levine, Richard Losick Biologia molecolare del gene, 8e 2022, pagine 944

Presentation of the course and its objectives. Methods of study and final examination. Cells as living entities and their properties. The structure of the prokaryotic cell. The eukaryotic cell: basic characteristics. The cell cycle and the anatomy of chromosomes. Mitosis and meiosis. Transmission of characteristics: Dominance and recessivity. Examples of genetic diseases. Macromolecules and their structures. History of the discovery of DNA from Mischer to Watson and Crick. The double helix of DNA and its chemical and physical properties. Spontaneous modifications of the bases and their effects on the conservation of genetic information. The secondary structures of DNA: DNA-B, DNA-A and DNA Z. The triple and quadruple helices of DNA. The role of cross-shaped structures in crossing over. The telomeres. DNA Tm and hyperchromism of the bases. The Cot value and complexity of the genome. The RNA world and its evolution. The secondary structures of RNA and their stability. The structural motifs of RNA, the double helix A of RNA. The different types of RNA and their structures. Ribozymes and their evolutionary and physiological significance as well as their applications in the biological field. Riboswitches and the role of RNA aptamers. The tertiary structure and topology of DNA. The quaternary structure of DNA: association with basic proteins. The influence of the bacterial chromosome. The influence of eukaryotic chromosomes: the role of histones. Chromatin remodelling and the role of histone tails. The dimensions of genomes and the paradoxes K, C and N. The role of repetitive sequences, pseudogenes and retropseudogenes. The retrotransposomes. Introns and their role. Epigenetic and its biological role. General principles of replication. The enzymes involved and the origins of replication. The phases of replication: initiation, elongation and termination. Bacterial chromosome replication Differences between prokaryotic and eukaryotic DNA polymerases. Comparison between prokaryote and eukaryote replication. Spontaneous changes in genomes and damage caused by external physical and chemical agents. Their effects on genomes. The repair systems, BERs and NERs, HEJ and NHEJ, of UV and mismatches. Translesion synthesis and direct couplings, transcription and DNA repair. General principles of transcription in prokaryotes. The initiation of transcription, evasion from the promoter and termination of transcription. The regulation of transcription by operons. Transcription in eukaryotes and its modulation. The general mechanism of splicing, autosplicing and trans-splicing. Alternative splicing and RNA editing. Translation and its general principles in prokaryotes and eukaryotes. The regulation of gene expression. The post-translational regulation of proteins. Recombinant DNA technology and cloning. The applications of recombinant DNA. The tools for cloning: Enzymes, vectors and hosts. Plasmids and phages as vectors. References to other vectors. Expression cloning. The steps of cloning and purification of recombinant proteins. Techniques to study genes and gene expression: direct and reverse hybridisation techniques, target amplification techniques (PCR, RT-PCR, qPCR and ddPCR), probe amplification techniques (LCR), signal (bDNA), isothermal amplification techniques (NASBA), gene sequencing techniques (Maxam-Gilbert-Sanger automation with electropherogram, pyrosequencing), epigenetic sequencing techniques (bisulfite).

Frontal teaching with the help of slides and videos. Involvement of students in exercises to link with other related disciplines (chemistry, biochemistry). Use of multimedia systems (interactive whiteboards, instant polling, wooclap) and class discussion. Involving students in exercises on using appropriate scientific language and solving simple biological problems (interactive whiteboards, instant polling, wooclap) and class discussion.

Teaching support with the availability of slides and educational materials on Teams

Oral exam: verification questions; relationship between the various processes of molecular biology. Open answers. Review questions with open-ended answers on the applications of molecular biology in the areas of the study of genes and gene expression in research and diagnostics. Assess ability to solve molecular biology problems in practical diagnostic applications. Ability to solve problems of molecular biology in a multidisciplinary team. Ability to critically discuss applications of molecular biology.

This course addresses issues closely related to one or more of the United Nations 2030 Agenda for Sustainable Development goals. In particular, it explains the evolution from the physiological to the pathological state of the human organism in relation to the environment in which it lives and lifestyle choices (primary prevention) and secondary prevention (early detection). It also explains the close connection between living beings for the well-being of ecosystems. Finally, it looks at biotechnologies used in medicine, agriculture, animal husbandry and industry, highlighting the strengths, weaknesses and risks.