The course aims to provide the fundamental knowledge of the molecular mechanisms within prokaryotic and eukaryotic cells that regulate and subtend the maintenance of genetic information and the flow of genetic information. During the course the main experiments useful for the achievement of current knowledge will be presented. Moreover, whenever possible, reference will be made to the application in the field of medical biotechnology of acquired knowledge. At the end of the course the students will have acquired a good knowledge of the main molecular biology techniques and the ability to make a correct one for the theoretical resolution of specific biological problems, remembering to set up the appropriate positive and negative controls in each experimental phase.
Expected learning outcomes
In-depth knowledge of the main mechanisms of molecular biology (replication, transcription and translation) and their regulation. Knowledge of the main molecular biology techniques useful in basic and applied research for the study of the molecular mechanisms underlying the correct flow of genetic information and its use.
Lesson period: First semester
(In case of multiple editions, please check the period, as it may vary)
Frontal teaching: Biological macromolecules: DNA, RNA, proteins (structure, function and purification techniques). Model organisms for molecular biology studies. Main molecular biology techniques for DNA, RNA and protein analysis and their manipulation (nucleic acid hybridization, Southern, Northern, in situ hybridization, DNA sequencing, PCR, SDS-PAGE, western blot, antibody production mono and polyclonal, HIC, Ip, ChIP, Transcriptome analysis, genome editing). DNA topology and topoisomerases. Chromatin condensation in eukaryotes: structure of nucleosomes and higher levels of condensation. Epigenetics: basic concepts (DNA methylation, post-translational modifications of histones and the hypothesis of the existence of an histone code). Brief review of transcription in prokaryotes: structure of the transcriptional apparatus, promoters and regulation mechanisms. The transcription cycle. The lactose and tryptophan operon. Transcription in eukaryotes: structure of transcriptional systems, promoters and regulation mechanisms. Particular emphasis will be given to class II transcription. Non-RNA coding: synthesis and function. RNA splicing: molecular mechanisms. Translation in prokaryotes and eukaryotes. DNA replication in prokaryotes and eukaryotes. Notes on the mechanisms of onset of mutations and DNA repair. Exercises: The boys will carry out a western blot and an immunofluorescence experiment in groups of two or maximum three students, discussing the expected results and those obtained at the end.
Prerequisites for admission
Students must have fulfilled all the prerequisite requirements indicated in the study plan: General and cellular biology, Genetics.
The course is subdivided into two parts. The first, representing the majority of the course, consists of frontal lectures, while in the second part the students (organized in small groups of two or three people) will independently carry out a laboratory activity by personally applying some of the molecular biology techniques explained in class. In all teaching moments, students are exhorted to try to find experimental strategies useful for solving small scientific problems or to critically analyze the results of some experiments. The teaching material consisting of presentations in PDF format is made available at the end of the lesson on the Ariel platform. Attendance of teaching is strongly recommended.
Students are provided with the pdf of the lessons taught. Each student must refer to a molecular biology textbook. In case of purchase, the following texts are recommended: MM Cox, Jennifer A. Doudna, MO'Donnel Molecular Biology principles and techniques. Zanichelli Watson JD, Baker TA, Bell SP, Gann A. Levine M. Losick Molecular biology of the Zanichelli gene. Amaldi et al. Molecular biology. Zanichelli
Assessment methods and Criteria
The level of learning is verified only at the end of the course, without any intermediate exam, through an oral interview. This is generally organized in two parts. In the first part, the student is asked two or three questions aimed at evaluating the knowledge of the main topics carried out in class (including also the laboratory experience). In this first phase the property of language, the ability to organize a linear discourse and, in general, the speaking capacity are also evaluated. The second part of the interview is generally proposed only to those students who have demonstrated a good familiarity with the subject. In this phase of the interview, a solution to a particular scientific problem is requested, using the molecular biology techniques explained in class. This phase of the interview, certainly more complex, serves to understand the student's ability to handle molecular biology and its methodological approaches by making diversified use of what has been learned in class. In this phase, it will be also evaluated the capacity of the student to organize a correct experiment, including the necessary controls and, when requested, the appropriate internal standards.