FA/0181 - GENOME VARIABILITY
Academic Year 2022/2023
Free text for the University
CARLA MARIA CALO' (Tit.)
- Teaching style
- Lingua Insegnamento
|[60/57] BIOLOGY||[57/20 - Ord. 2018] Bio-Molecolare||7||60|
The course aims to provide a basic knowledge of the molecular mechanisms related to the maintenance and variability of genetic information in prokaryotes and eukaryotes. This objective will be achieved by integrating knowledge of microbiology, molecular biology, genetics and population genetics.
At the end of the course the student must know the extent and importance of the variability of human DNA and how to measure it. He will have to be able to elaborate and interpret autonomously the genetic data obtained both experimentally (genotyping) and theoretically (bibliographic research); he must also possess a basic knowledge of the IT tools used for DNA analysis.
At the end of the course the student will also have acquired basic notions on recombinant DNA methodologies and related biotechnological applications, on the methods used for massive parallel DNA sequencing and gene expression studies.
KNOWLEDGE AND UNDERSTANDING ABILITY.
Theoretical lessons aimed at providing the student with in-depth knowledge of the structure, replication, maintenance of integrity and the variability of the genome of microorganisms, genetic variability and polymorphisms of human DNA and their application in the population field, basic knowledge on technologies used for the isolation, study and manipulation of genes and the methods used for the study and evaluation of gene expression.
Practical and autonomous guided activities for the research and comparison of DNA sequences and mutated sites:
- computer lab work for the research of scientific publications;
- individual work in the computer room for the search for sequences and the comparison between them, in order to extract the necessary information.
Acquisition of adequate knowledge on the various applicative possibilities of biotechnologies in the biomedical field, in agriculture and in the pharmaceutical industry
AUTONOMY OF JUDGMENT
- Acquisition of the ability to autonomously process and interpret genetic data related to sequence mutations and generally to the variability of human DNA
- Acquisition of the evolutionary importance of the variability of human DNA;
- Acquisition by the student of the correct evaluation of the use of the most suitable biotechnologies based on the different applications
ABILITY IN COMMUNICATION
Acquisition of the ability to fully and clearly illustrate the laws that govern the genetic variability of microorganisms and humans. Ability to use an appropriate scientific language
Acquisition of the ability to illustrate biotechnologies and their applications in a complete and clear way
CAPACITY TO LEARN
Acquisition of adequate scientific knowledge of genetic variability e
of his study methods, through lectures and specific scientific texts of
in-depth study of the Microbiology and Biology of Human Populations or Molecular Anthropology. Consultation of databases and online bibliographic research, basic cognitive tools necessary for the continuous updating of knowledge.
Acquisition of adequate scientific knowledge on the various application possibilities of biotechnology through lectures, multimedia activities and the ability to perform bibliographic on-line searches that allow continuous updating on these topics
Basic knowledge of genetics and / or molecular biology, in particular on the structure of DNA
Module1 (2CFU): (Mechanisms of) variability of the genome of microorganisms
DNA structure. Topological DNA changes and DNA topoisomerase (3h)
DNA replication. Bacteria, Archea, Eukarya DNA polymerases (3h)
Recognition of the origin of replication in bacteria and Eukarya. Replicative strategies of linear genomes (2h) Mismatch repair. DNA damage: spontaneous damage and mutagens (chemical and physical). Assays for the evaluation of genotoxic agents in bacteria and eukaryotes. Shelter systems. Direct shelter. Shelter for excision: BER, NER, RER. (4h)
Recombination mechanisms. Homologous recombination (Locus MAT in Saccharomyces), site specific recombination, transpositional recombination. (4h)
Module 2 (2 + 1 CFU): Variability of human DNA
Variability of human DNA. What is meant and measured. Human variability as a mechanism of survival of the species. (2h)
The satellite DNA: STRs and VNTRs. origin, description, mutation rates and applications. Copy Number Variation: definition and applications. (4h)
SINE and LINE polymorphisms. Deepening on ALUs. Origin and applications. (2h)
SNPs: mutation rates, utilities, identification methods. The restriction enzymes for the recognition of SNPs. (2h)
Uniparental transmission markers: mtDNA, advantages and applications, the non-recombinant portion of the Y chromosome: polymorphisms used and applications, haplotypes and haplogroups (4 hours)
The fallacy of the biological concept of the human race according to molecular studies. (2h)
Laboratory: Human DNA extraction. Methods for detecting polymorphism: indels and SNPs. PCR, enzymatic digestion and electrophoresis.
Module 3 (2CFU): recombinant DNA and biotechnology
Recombinant DNA technologies: vectors used for bacterial cloning, in yeast and for genetic transfer in animal and plant cells. Genoteche, strategies of gene cloning in prokaryotic cells, in yeast, in animal and plant cells (5h)
Biotechnological applications of cloning: production of recombinant proteins in E.Coli, in insect cells. Production of recombinant proteins with applications in the biomedical field. Gene transfer for plant genetic manipulation Problems associated with the use of transgenic plants (5h)
New generation sequencing techniques (NGS): NGS sequencing platforms. Library preparation, genome sequencing, target region sequencing. Data analysis. The Human Genome Project, comparative genomics. (3h)
Analysis methodologies for the study of gene expression: general aspects. RT-PCR, in vitro mutagenesis, RNA interference assays (RNAi), reporter genes. Study of the gene expression profile by microarray. Transcriptome analysis using NGS technologies (RNA-Seq) (3h).
Teaching will be organized in frontal classes, integrated and "augmented" with online strategies, in order to guarantee its use in an innovative and inclusive way.
The credits for the frontal lessons are 4, for a total of 32 hours. The lessons will be in Italian.
The laboratory credits (1 CFU for a total of 12 hours) will take place in the computer room, where students will learn and deepen research methods in specialized databases and align and compare two or more DNA or RNA sequences .
Verification of learning
The final exam will be oral. The contents will be evaluated, but also the ability to analyze and summarize and the correctness of the scientific language used.
The final grade takes into account various factors:
Quality of knowledge, skills, skills possessed and / or manifested:
a) appropriateness, correctness and congruence of knowledge
b) appropriateness, correctness and congruity of skills
c) appropriateness, correctness and congruity of competences
a) Expressive capacity;
b) appropriate use of the specific language of the discipline;
c) Logical abilities and consequentiality in the connection of the contents;
e) Synthesis ability also through the use of the proper symbolism of the matter and the graphic expression of notions and concepts, in the form for example of formulas, schemes, equations.
Dehò & Galli. Biologia dei microrganismi, Ambrosiana 2017.
Lewin Il Gene X, Zanichelli, 2012
Relethford John H. Genetica delle popolazioni umane. Ed. Ambrosiana, 2013
Brown Terry A. Biotecnologie molecolari. Principi e Tecniche Ed Zanichelli, 2017
Materials provided by the teachers.
Main slides of the course and scientific articles to supplement the textbook will be available for the students.