The course aims to make the student understand some important biological processes through the combination of two points of view, the biochemical-structural and the genetic-informational.

In this way we will try to provide the student with the cultural bases to understand how the genome is structured and how it is organized, and the molecular and cellular mechanisms that underlie genome expression.

Some of the main techniques used in molecular biology, as they are applied, will be discussed 

Nucleic acid structure, DNA topology and DNA topoisomerase. RNA structure.

Genetic code, Reading phases and ORF. Suppression mutations and genetic code. Chromosomes and chromatin. Organization and packaging of eukaryotic DNA. Centromeres, Telomeres, Nucleosomes and structural and functional properties of chromatin. DNA replication. Nucleosomes and transcription of genes, Post-translational modifications of histones and modulation of chromatin transcriptional activity.

Prokaryotic and eukaryotic genomes. Role of introns. Pseudogenes. Repeated sequences of eukaryotic genomes. Satellite or highly repetitive DNA. Microsatellites and minisatellites. Repetitive DNA sequences interspersed in the genome. Mitochondrial genome and disorders. Virus genome.

REPLICATION AND MAINTENANCE OF THE GENOME. Replication in eukaryotes. Replication of telomeres in the linear chromosomes of eukaryotes. Control of DNA replication during the cell cycle.

Alterations in replication mechanisms. DNA repair systems (NER, MMR, PRR). Cellular response to damage to DNA and connections between repair and cell cycle. Recombination. Recombination in meiosis, homologous or generalized recombination. 

Properties and viral cycle of lambda phage. Transposition. The retroviruses. Rearrangements of DNA sequences and control of gene expression. Transcription and regulation in prokaryotes.

Transcription and regulation in eukaryotes. 

Protein Synthesis in prokaryotes, in eukaryotes, lengthening. Protein synthesis inhibitors and antibiotics.

Translation adjustment. Regulation of stability and degradation of mRNAs. Other levels of regulation: epigenetic and post-translational regulation, non-coding RNA roles (ncRNA). Epigenetic adjustments: DNA methylation and chromatin remodeling. DNA methylation and gene expression. Identification of methylated CpGs in genomic DNA. Genetic imprinting Epigenetic regulation and pathology. Chromatin remodeling.

Regulatory roles of ncRNAs. sRNA, MicroRNA, Long non-coding RNA (lncRNA). Post-translational modifications and adjustments of proteins. Protein Ubiquitination, Protein Sumoylation, Phosphorylation and Protein Dephosphorylation. Protein acetylation. Protein methylation. Glycosylation and lipid modifications of proteins.

Molecular Biology Techniques: spectrophotometry for DNA denaturation and reassociation analysis, Absorption spectrum. DNA denaturation, Tm. Use of DNA probes. DNA marking with radioactive atoms. In situ or cytological hybridization, Southern and Northern blot, Colony hybridization or on plaque, Microarray (or microchip). Ultracentrifugation (sedimentation in sucrose gradients, density gradients of cesium chloride)

Restriction enzymes and DNA ligases. Cloning vectors. Genomic DNA banks. Genomic DNA library. cDNA libraries. PCR. Determination of DNA nucleotide sequence. New generation sequencing platforms. Inhibition of the expression of specific genes: antisense RNA and RNA interference. Bioinformatics and Genomics: Sequencing and assembly of complete genomes, Transcriptome sequencing, Identification and annotation of genes. Comparison and alignment of sequences. 

At the end of the course the student must have the tools to understand the terminology used in the field of molecular biology, the molecular basis of biological systems and processes, the mechanisms and the different levels of control of gene expression, the basic techniques for the study of nucleic acids.

The teaching material prepared by the lecturer in addition to recommended textbooks (such as for instance slides, lecture notes, exercises, bibliography) and communications from the lecturer specific to the course can be found inside the Moodle platform › blended.uniurb.it

Assessment

Multiple choice test and oral exam.

Disabilità e DSA

Le studentesse e gli studenti che hanno registrato la certificazione di disabilità o la certificazione di DSA presso l'Ufficio Inclusione e diritto allo studio, possono chiedere di utilizzare le mappe concettuali (per parole chiave) durante la prova di esame.

A tal fine, è necessario inviare le mappe, due settimane prima dell’appello di esame, alla o al docente del corso, che ne verificherà la coerenza con le indicazioni delle linee guida di ateneo e potrà chiederne la modifica.