Activity | Credits | Period | Academic staff | Timetable |
---|---|---|---|---|
teoria | 9 | II semestre | Daniele Guardavaccaro | |
laboratorio [laboratorio 1° turno] | 3 | II semestre | Daniele Guardavaccaro | |
laboratorio [laboratorio 2° turno] | 3 | II semestre | Massimiliano Perduca |
The main goal of this course is to provide the students with the basic knowledge of the composition, structure and interactions of cellular molecules, such as nucleic acids and proteins, that carry out essential biological processes in eukaryotic and prokaryotic cells.
THEORY
DNA AND CHROMSOMES
The structure and function of DNA
Chromosomal DNA and its packaging in the chromatin fiber
Chromatin structure and function
The global structure of chromosomes
DNA REPLICATION, REPAIR AND RECOMBINATION
DNA replication mechanisms
The initiation and completion of DNA replication in chromosomes
DNA repair
Homologous recombination
Transposition and conservative site-specific recombination
HOW CELLS READ THE GENOME: FROM DNA TO PROTEIN
From DNA to RNA
From RNA to protein
Protein degradation and the ubiquitin-proteasome system
CONTROL OF GENE EXPRESSION
Control of transcription by sequence specific DNA-binding proteins
Transcription regulators
Molecular genetic mechanisms that create and maintain specialized cell types
Mechanisms reinforcing cell memory in plants and animals
Post-transcriptional controls
Regulation of gene expression by non-coding RNAs
ANALYZING CELLS AND MOLECULES
Isolating cells and growing them in culture
Analyzing and manipulating DNA
Studying gene expression and function
MEMBRANE STRUCTURE
The lipid bilayer
Membrane proteins
TRANSPORT OF SMALL MOLECULES ACROSS MEMBRANES
Principles of membrane transport
Transporters and active membrane transport
INTRACELLULAR COMPARTMENTS AND PROTEINS SORTING
The compartmentalization of cells
The transport of molecules between the nucleus and cytosol
The endoplasmic reticulum
Molecular mechanisms of protein translocation across the ER membrane
INTRACELLULAR MEMBRANE TRAFFIC
Molecular mechanisms of membrane transport
Transport from the ER through the Golgi apparatus
Transport from the trans-Golgi network to lysosomes
Transport into the cell from the plasma membrane: endocytosis
Transport from the trans-Golgi network to lysosomes
Transport from the trans-Golgi network to the cell exterior: exocytosis
CELL SIGNALING
Principles of cell signaling
Signaling through G-protein-coupled receptors
Signaling through enzyme-coupled receptors
Alternative signalling routes in gene regulation
THE CYTOSKELETON
Function of the cytoskeleton
Actin and actin-binding proteins
Myosin and actin
Microtubules
Intermediate filaments and septins
Cell polarization and migration
THE CELL CYCLE
Overview of the cell cycle
The cell cycle control system
S-phase
Mitosis
Cytokinesis
Meiosis
Control of cell division and cell growth
CELL JUNCTIONS AND THE EXTRACELLULAR MATRIX
Cell-cell junctions
The extracellular matrix of animals
Cell-matrix junctions
CANCER
Cancer as a microevolutionary process
Cancer-critical genes: oncogenes and tumor suppressors
Cancer therapy
One credit of the course (corresponding to 8 hours) will be kept for the students to discuss a topic chosen from the research literature in molecular biology.
PRACTICAL:
Subcloning and protein expression:
PCR
Electrophoresis and DNA gel extraction
Plasmid DNA purification
Digestion with restriction enzymes and ligation
Transformation and colony PCR
Heterologous expression of a recombinant protein in E. coli and analysis by SDS-PAGE and Western Blotting.
Oral examination. The final exam consists of three questions concerning any of the topics treated during the course. It is passed if all answers are positive.
Activity | Author | Title | Publisher | Year | ISBN | Note |
teoria | Jocelyn E. Krebs, Elliott S. Goldstein, Stephen T. Kilpatrick | Lewin's Genes XII (Edizione 12) | Jones & Bartlett Pub | 2017 | 1284104494 | |
teoria | Bruce Alberts, Alexander Johnson, Julian Lewis, David Morgan, Martin Raff | Molecular Biology of the Cell (Edizione 7) | Garland Science | 2017 | 0815344643 | |
teoria | Nancy Craig, Rachel Green, Carol Greider, Gisela Storz, Cynthia Wolberger, Orna Cohen-Fix | Molecular Biology: Principles of Genome Function (Edizione 2) | OUP Oxford | 2014 | 0199658579 | |
teoria | Harvey Lodish, Arnold Berk, Chris A. Kaiser, Monty Krieger, Anthony Bretscher, Hidde Ploegh, Angelika Amon, Kelsey C. Martin | Molecular Cell Biology (Edizione 8) | Macmillan | 2016 | 9781464187445 | |
teoria | Erik Pierre | Molecular Cloning (Edizione 1) | Ml Books International | 2015 | 1632394685 | |
teoria | Michael R. Green e Joseph Sambrook | Molecular Cloning: A Laboratory Manual, Fourth Edition (Edizione 4) | CSHL Press | 2012 | 978-1-936113-42-2 | |
teoria | Geoffrey M. Cooper, Robert E. Hausman | The cell: a molecular approach (Edizione 6) | Sinauer Associates, Inc | 2013 | 978-1-60535-155-1 |
Title | Format (Language, Size, Publication date) |
01 PCR di amplificazione del frammento da Miniprep |
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02 Preparazione gel di agarosio 0.8% per la purificazione dell’amplificato |
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03 Purificazione dell’inserto da gel utilizzando il GenElute™ Gel Extraction Kit |
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04 Clonaggio del cDNA di una proteina umana in vettore pET22-b |
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05 Ligazione dell’inserto nel vettore pET22-b |
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06 Preparazione delle piastre di agar per XL1-Blue |
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07a Preparazione delle cellule competenti XL1Blue |
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07b Scheda preparazione delle cellule competenti XL1Blue |
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08 Trasformazione delle cellule di di E. coli XL1Blue |
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09 Colony PCR e piastre replica |
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10a Miniprep Sigma |
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10b Tagli enzimatici di controllo |
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11 Trasformazione ceppo BL21 |
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12 Induzione ceppo BL21 |
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13 SDS PAGE |
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14 Western Blot |
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I-1 Ballas et al |
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I-2 Busino et al |
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I-3 Carrano et al |
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I-4 Cepeda et al |
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I-5 Duan et al |
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I-6 Duan2 et al |
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I-7 Jin et al |
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I-8 Koo et al |
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II-1 Lee et al |
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II-2 Mamely et al |
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II-3 Mohamed et al |
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II-4 Nakano et al |
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II-5 Raducu et al |
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II-6 Strohmaier et al AND Koepp et al |
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II-7 Tomás-Loba et al |
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II-8 Zhang et al |
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Laboratorio Didattico Biologia Molecolare II Turno |
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Laboratorio Didattico Biologia Molecolare I Turno |
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Sequenza genica e primers |
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