The purpose of this course is to provide students with the necessary information to understand the theoretical foundations of classical and molecular genetics and the experimental approaches that have allowed its definition. Students will be expected to learn the logic of formal genetic analysis and the methodologies of genetic dissection of biological phenomena. Concepts for understanding the paradigm shift that has occurred in the post-genomic era will be provided, and an understanding of the importance of both eukaryotic and prokaryotic model systems will be stimulated. The course is intended to provide the basis for further in-depth study in all areas of genetics.
EXPECTED LEARNING OUTCOMES 1) Knowledge and understanding: At the end of the training activities, students will be able to manage in a project-oriented way the complementary methods derived from the knowledge of recombinant DNA techniques. 2) Ability to apply knowledge and understanding : Students will be stimulated to select the most appropriate and relevant methodological/experimental approaches to achieve specific objectives (e.g. biotechnological production of drugs, recombinant proteins, etc.). 3) Making judgements: The teaching will provide the student with the ability to work independently by providing appropriate types of teaching materials (lectures in the form of presentations, specific monographs, relevant scientific literature, computer platforms) and the performance of congruent laboratory activities synchronized with the theoretical part of the course. 4) Communication skills: students will be encouraged to actively participate in the lessons and will be stimulated to design and solve specific scenarios related to the different evolutionary phases of the course. 5) Learning skills: The students' learning skills will be assessed in itinere and verified through the individual ability to solve relevant and specific scenarios of interest, different from those envisaged during the course.
Genetics program (9CFU); Undergraduate Course of Biotechnology (L-2), 2022-2023 Professor: Pasquale Mosesso 1. Mendelian inheritance and chromosome theory of inheritance Mendel’s postulates and elements of mendelian genetics applied to man and agriculture. Laws of probability and their relevance to explain genetic events (the product law and the sum law); Use of chi-square analysis to evaluate the influence of chance on genetic data, interpretation of chi-square calculations. Cell cycle; mitosis, meiosis and concordance between meiosis and genetic mendelism. Demonstration of chromosome theory of inheritance. Chromosomal sex-determining system; sex-limited and sex-influenced inheritance. 2. Extensions of mendelian genetics Alleles alters phenotypes in different ways: Incomplete dominance, codominance, multiple alleles, lethal alleles. Environment and gene expression: Effects of “internal” and “external” environments. Penetrance and expressivity. Gene interaction. Epistasis and pleiotropy. 3. Genes linkage and chromosome mapping The crossing -over. Linkage maps: Mapping with three-point crosses; Interference; Some examples of linkage maps; Chi-squared test. 4. Molecular structure and replication of genetic material Molecular structure of DNA and RNA: Identification of DNA as genetic material (experiments of Hershey and Chase). Characteristics of nucleic acids (experiments of Chargaff). Replication of DNA in the prokaryotes and eukaryotes. Demonstration that reproduction of DNA is by semiconservative replication (experiments of Meselson e Stahl). 5. Genetic analysis in bacteria The bacterial chromosome: Transfer of genetic material in bacteria: Transformation; Conjugation; sexduction and transduction. Methods of genetic mapping in bacteria and bacteriophages. Extrachromosomal elements: Natural and artificial plasmids. 6. Molecular properties of genes Structure and function of genes: Components of the eukaryotic chromosomes (DNA, histonic and non-histonic protein); Chromatin structure and nucleosomes; Repetitive DNA sequences and DNA satellite; How genes work (hypothesis one gene one protein); colinearity between gene and protein. Complementation and mutation sites. Translation of mRNA; Structure and functions of tRNA. The genetic code; Studies by Nirenberg, Matthaei and Others led to the deciphering of the code; Early studies established the basic operational pattern of genetic code: The triplet nature of the code; the nonoverlapping nature of the code; the commaless and degenerate nature of the code. 7. Mechanisms of production of genetic variability Definition and classification of mutations: Gene mutations: base pair substitutions, insertions, deletions; reversion and suppression of mutations; phenotypic effects of gene mutations. Fluctuation test and spontaneous mutation in bacteria (neo-Darwinian theory). Chromosome mutations: structural (chromosomal aberrations) and numerical (aneuploidy, polyploidy). Environmental agents which damage DNA: Ionizing electromagnetic radiations (X-rays, gamma-rays, betatron-synchrotron radiations); Ionizing subatomic particles (alpha particles, beta particles neutrons). Non-ionizing electromagnetic radiations (UV-A, UV-B, UV-C); UV and ozone. Electromagnetic fields (microwaves, HF, ELF etc.). Chemical mutagens: alkylating agents; environmental pollutants in the cities [car exhaust, sulphur and nitrogen oxides, polycyclic aromatic hydrocarbons (PAH), formaldehyde, asbestos]; food additives and contaminant of aliments (aflatoxins, ochratoxins nitrosamines, heterocyclic aromatic amines, phenols of vegetal origin, etc); therapeutic agents (mitomycin-C, bleomycin, nitrogen mustards, inhibitors of DNA topoisomerases I and II, cytostatic agents); pesticides. Biological mutagens: microbial and viral pathogens; transposable elements. DNA repair systems: fotoliase, alchiltransferases, “nucleotide excision repair” (NER), “base excision repair (BER), “mismatch repair” (MMR), “SOS” repair, recombinational repair: repair of DNA single strand breaks (SSB) and DNA double strand breaks (HR e NHEJR); DNA damage, activation of cellular “checkpoints. Transposable elements: Bacterial and eukaryotic transposable elements; Retrovirus; Transposons and chromosome mutability. 8. Gene regulation and expression in bacteria Negative and positive regulation; Genetic dissection of lac operon in E. coli; The operon of tryptophan. 9. Gene regulation and expression in eukaryotes Gene expression regulation at epigenetic, transcriptional and post-transcriptional levels. 10. Principles of Genetic engineering Basic techniques to manipulate genetic material (DNA and RNA): Restriction enzymes; Cloning of DNA fragments. Chromosomal and plasmidico DNA separation and purification. Techniques of analysis of DNA, RNA: Gel Electrophoresis; Amplification of nucleic acid fragments by Polymerase Chain Reaction; Hybridization, Southern Blotting, and Northern Blotting. DNA sequencing methods: General principles of the procedure; sequencing of large genomic regions. In vitro mutagenesis. 11. Population genetics analysis Allele frequencies in populations vary in space and time. The Hardy-Weinberg law describes the relationship between allele frequencies and genotype frequencies in an ideal population. The Hardy-Weinberg law can be used in human populations for multiple alleles, X-linked traits and estimating heterozygote frequencies. Natural selection is a major force driving allele frequency change: Natural selection, fitness and selection, selection in natural populations. Mutation creates new alleles in a gene pool. Migration and gene flow can alter frequencies. Genetic drift causes random changes in allele frequencies in small populations. Non-random mating changes genotype frequency but not allele frequency: Inbreeding; genetic effects of inbreeding.