MOLECULAR BIOLOGY
- Overview
- Assessment methods
- Learning objectives
- Contents
- Full programme
- Delivery method
- Teaching methods
- Contacts/Info
Although no preparatory skills are specifically required, knowledge of Biochemistry and the general concepts of Cell Biology and Genetics are fundamental to tackle the course and understand the covered topics.
Verification of learning takes place through a written test and is aimed at assessing the achievement of learning outcomes and therefore the level of knowledge and understanding of the theoretical bases of Molecular Biology and the ability to present the topics covered in the course, using the appropriate scientific terminology. The exam will consist of 4 open questions and 6 multiple choice questions, which will span the entire program. The accuracy and completeness of the information presented in response to the open questions will weigh 90% on the final evaluation. The exam result will be out of thirty. The test is considered passed with a vote of at least 18/30. The criteria with which the verification of the acquired knowledge and skills will be carried out are: the degree of in-depth exposition of the subject matter of the question, the critical ability to connect and elaborate the knowledges, the clarity of the concepts presented and the propriety of the terminology used. Candidates who have scored 24 or more in the written test may opt for an oral test aimed at improving their grade.
The practical activities will be evaluated by an oral exposition of the obtained results. Each group of students who carried out the same set of experiments will be asked to prepare a PowerPoint presentation reporting about procedures and the experimental outcomes, which they will then discuss with the lecturer and their colleagues. The final mark will be the arithmetic average of the outcome of the two tests (the written exam and the oral exposition of the practical activities), each weighed for the relative ECTS.
Educational objectives
The lecturer with this course aims to provide an in-depth knowledge of biological systems at the molecular level. In particular, lectures will deal with the processes through which gene information is translated into protein products and the different mechanisms involved in their regulation. The main objective is to make students acquire a solid preparation for understanding the molecular basis of these processes and then, continuing along their studies, the more advanced aspects of their biotechnological applications. In this perspective, the course will also allow students to learn and practice the main techniques currently applied in research: through the practical activities in the laboratory, they will be able to organize and set up the experiments, work in groups and evaluate and in-terpret the experimental data.
The course is part of the II year (II semester). It will allow the student to integrate the notions acquired in the genetics and biochemistry courses and to understand how to apply them to the modification of biological systems, so as to be able to contribute to research projects and / or application processes in the biotechnological field, in line with the objectives of the CdS.
Learning Outcomes
At the end of the course, the student will be able to:
- explain the molecular details of DNA replication, transcription and translation processes and their role at the cellular level;
- understand and discuss the role of the genetic code as a universal information management system in living beings and the mechanisms for regulating gene expression;
- know and explain the consequences of alterations and defects in the processes treated and the mechanisms involved in their resolution;
- apply the concepts learned to DNA manipulation techniques within a laboratory and in the field of biotechnology research;
- communicate with appropriate scientific terminology.
The topics covered during the course are the following:
Brief introduction to biological macromolecules
The condensation of DNA in the different biological systems
Molecular Biology Techniques
DNA replication
Mutations, base modifications and chromosomal alterations
DNA Repair Systems
Recombination
Transcription
Eukaryotic RNA processing and maturation
Protein synthesis
Transcription regulation
Genomic editing
During the practical activities in the laboratory, students will deal with the cloning of a coding sequence into an expression vector, the transformation of an E. coli strain, the extraction of plasmid DNA and the restriction analysis to verify the success of the cloning.
The topics covered during the course are the following:
Brief introduction to biological macromolecules
- DNA topology
The condensation of DNA in the different biological systems
- brief hints on the containment strategies of genomes from viruses to eukaryotes
- nucleosomes and chromatin
Molecular Biology Techniques
- preparation, analysis and manipulation of nucleic acids
- cloning and vectors for cloning and expression, transformation and selection
- PCR, basic principles, RT-PCR, sequencing
- Hybridization and genomic library analysis
DNA replication
- the origins of replication and the replicon model
- DNA polymerase (catalyzed reactions, accuracy and fidelity)
- the replication fork and the replisome complex
- elongation, termination and their regulation
Mutations, base modifications and chromosomal alterations
DNA Repair Systems
- mismatch repair, direct repair of damage, by excision of bases (BER) or nucleotides (NER)
- mechanisms of tolerance
Recombination
- homologous recombination and the role in DNA lesion repair
- site specific recombination
- transposition
Transcription
- molecular mechanisms in prokaryotes (concept of operon, structure / function of RNA polymerase, promoters and their recognition, stages of the process)
- molecular mechanisms of eukaryotes (RNA polymerases I, II and III, specific promoters, process steps)
Eukaryotic RNA processing and maturation
- capping, polyadenylation and splicing of introns in mRNAs
- hints to post-transcriptional modifications of rRNA and tRNA
- RNA editing; transport and stability
Protein synthesis
- tRNA, aminoacyl-tRNA synthetase, ribosomes and ribosomal proteins
- the molecular mechanisms (initiation, elongation, translocation and termination) and the factors involved
- structure, ownership and use of the genetic code; Codon-anticodon interaction
Transcription regulation
- molecular mechanisms in prokaryotes: role of regulatory proteins
-examples: regulation of lac operons, trp, araBAD, the SOS response
- molecular mechanisms in eukaryotes: the role of transcriptional activators
-signal transduction, gene silencing and epigenetic control
Small regulatory RNA
- CRISPR sequences in bacteria
-synthesis and function of miRNAs in eukaryotes
Genomic editing
- ZFN and TALEN
- the CRISPR / Cas9 system
During the practical activities in the laboratory, students will deal with the cloning of a coding sequence into an expression vector, the transformation of an E. coli strain, the extraction of plasmid DNA and the restriction analysis to verify the success of the cloning.
The course includes lectures: the discussion of the various topics will be carried out through Power-Point presentations (with slides in English). Videos will be made available among the didactic material to help understand some of the processes and macromolecular systems involved. Students will be suggested a number of text books to choose from to complete their preparation. To allow students self-assessment of the level of learning, at the end of each macrotopic a series of multiple choice questions are proposed in the classroom, which are then discussed in the classroom (these tests are not withdrawn by the teacher and have no weight in the classroom. final evaluation).
The lecturer is always available to receive students, preferably by appointment (via requests to the email address silvia.sacchi@uninsubria.it). Prof. Sacchi is also available for in-depth or clarification meetings for groups of students on the topics covered by the course, that will be scheduled in the same way.