With this course students will acquire the fundamental knowledge of the molecular mechanisms regulating the maintenance of genetic information and its decoding. Both prokaryotic and eukaryotic cells will be considered.
The course aims to provide the knowledge of the main techniques of molecular biology and the ability to properly use them for the theoretical resolution of specific biological problems, remembering to set up in each experimental phase the appropriate positive and negative controls.
Lectures: Biological macromolecules: DNA, RNA, proteins (structure, function and purification techniques). Model organisms for molecular biology studies. Major molecular biology techniques for the analysis of DNA, RNA and proteins and their manipulation (hybridization of nucleic acids, Southern, Northern, in situ hybridization, DNA sequencing, PCR, qPCR, SDS-PAGE, Western blot , production of monoclonal and polyclonal antibodies, HIC, Ip, chIP, a mention of the transcriptome). DNA topology and topoisomerases. Chromatin condensation: nucleosome structure and higher levels of condensation. Epigenetics: basic concepts. Brief review of transcription in prokaryotes: transcriptional apparatus; structure of the promoters and regulatory mechanisms. The cycle of transcription. Transcription in eukaryotes: structure of the transcriptional apparatus, promoters and regulatory mechanisms. Non coding RNAs: synthesis and function. RNA splicing: molecular mechanisms. Translation in prokaryotes and eukaryotes. DNA replication in prokaryotes and eukaryotes.
Exercises: Groups of 4-5 students will be provided with experimental projects to be developed using the knowledge acquired during the course. At the end, each group will prepare a manuscript containing a brief introduction of the scientific problem followed by a detailed description of the chosen experimental approaches, the techniques used and the corresponding controls.