Alessandra TOSCO | MOLECULAR BIOTECHNOLOGY
Alessandra TOSCO MOLECULAR BIOTECHNOLOGY
cod. 0760300033
MOLECULAR BIOTECHNOLOGY
| 0760300033 | |
| DIPARTIMENTO DI FARMACIA | |
| EQF7 | |
| PHARMACY | |
| 2023/2024 |
| YEAR OF COURSE 4 | |
| YEAR OF DIDACTIC SYSTEM 2023 | |
| AUTUMN SEMESTER |
| SSD | CFU | HOURS | ACTIVITY | |
|---|---|---|---|---|
| BIO/11 | 6 | 48 | LESSONS |
| Objectives | |
|---|---|
| THE COURSE AIMS TO PROVIDE INFORMATION ON THE MAIN MOLECULAR BIOLOGY TECHNIQUES (CLASSICAL AND INNOVATIVE) USED FOR THE PREPARATION, ANALYSIS AND MANIPULATION OF NUCLEIC ACIDS AND PROTEINS. KNOWLEDGE AND UNDERSTANDING: AT THE END OF THE COURSE THE STUDENT WILL KNOW THE MAIN TECHNIQUES FOR: -DNA AND RNA ISOLATION AND ANALYSIS -PRODUCTION OF GENETICALLY MODIFIED ORGANISMS -THE PRODUCTION OF BIOTECHNOLOGICAL DRUGS -PROTEIN-PROTEIN AND PROTEIN-DNA INTERACTIONS ABILITY TO APPLY KNOWLEDGE AND UNDERSTANDING: THE STUDENT APPLYING THE ACQUIRED SKILLS MUST BE ABLE TO INDICATE THE APPROPRIATE METHODOLOGIES TO BE APPLIED TO BIOLOGICAL RESEARCH PROBLEMS IN THE CONTEXT OF: -GENE EXPRESSION ANALYSIS -PRODUCTION OF GENETICALLY MODIFIED ORGANISMS -PRODUCTION OF BIOTECHNOLOGICAL DRUGS -PROTEIN-PROTEIN AND PROTEIN-DNA INTERACTIONS |
| Prerequisites | |
|---|---|
| THE PREREQUISITE OF MOLECULAR BIOLOGY IS REQUIRED. |
| Contents | |
|---|---|
| -DNA AND RNA ANALYSIS: PURIFICATION OF GENOMIC AND PLASMID DNA AND RNA; QUANTITATIVE AND QUALITATIVE ANALYSIS: SPECTROPHOTOMETRY, ELECTROPHORESIS AND TRANSFER METHODS (4 HOURS OF THEORETICAL LESSONS). -ENZYMES OF RESTRICTION AND MODIFICATION, TRANSFORMATION AND TRANSFECTION, CLONING VECTORS: PLASMIDS, LAMBDA, M13 AND COSMIC VECTORS; ARTIFICIAL CHROMOSOMES: YAC, PAC, BAC AND HAC (6 HOURS OF THEORETICAL LESSONS). -THE PRODUCTION OF RECOMBINANT PROTEINS: EXPRESSION IN E. COLI; EXPRESSION VECTORS; CONSTITUTIVE AND INDUCIBLE PROMOTERS; YEAST EXPRESSION: S. CEREVISIAE, P. PASTORIS, EXPRESSION IN INSECT CELLS AND IN HIGHER EUKARYOTIC CELLS. THE USE OF RECOMBINANT PROTEINS IN SCIENTIFIC RESEARCH AND IN THE PHARMACEUTICAL INDUSTRY, BIOTECHNOLOGICAL DRUGS, THE PRODUCTION OF MONOCLONAL ANTIBODIES (8 HOURS OF THEORETICAL LESSONS). -PCR AND ITS APPLICATIONS IN THE BIOMEDICAL FIELD: REACTION CONDITIONS; QUANTITATIVE PCR; PCR IN THE DIAGNOSIS OF GENETIC DISEASES; PATERNITY TESTING; FORENSIC APPLICATIONS (4 HOURS OF THEORETICAL LESSONS). -DNA SEQUENCING: THE BASIC TECHNIQUES FOR SEQUENCING; GENOME SEQUENCING; THE HUMAN GENOME PROJECT. GENE CLONING STRATEGIES: GENOMIC DNA AND CDNA LIBRARIES; ANALYSIS OF A GENOTHEQUE; HYBRIDIZATION, CHROMOSOME WALKING AND DIFFERENTIAL ANALYSIS (4 HOURS OF THEORETICAL LESSONS). -ANALYSIS OF A PROMOTER: CONSTRUCTION OF REPORTER PLASMIDS; ELECTROPHORETIC MOBILITY TEST; FOOTPRINTING; CHROMATIN IMMUNOPRECIPITATION (4 HOURS OF THEORETICAL LESSONS). -STUDY OF PROTEIN-PROTEIN INTERACTIONS: THE DOUBLE HYBRID; CO-IMMUNOPRECIPITATION; "FISHING FOR PARTNERS"; ANALYSIS USING SPR TECHNOLOGY (2 HOURS OF THEORETICAL LESSONS). -GENE TRANSFER INTO ANIMAL CELLS: CELL CULTURES; TRANSFECTION TECHNIQUES. GENETIC MANIPULATION OF ANIMALS: INJECTION INTO THE PRONUCLEUS; EMBRYONIC STEM CELLS; METHODS FOR THE PRODUCTION OF TRANSGENIC MICE; DNA TRANSFER TO OTHER VERTEBRATES; GENE THERAPY. POST-GENOMIC ANALYSIS: GLOBAL CHANGES IN GENE EXPRESSION; MICROARRAY; PROTEOMIC ANALYSIS (4 HOURS OF THEORETICAL LESSONS). LABORATORY EXERCISES: -PREPARATION OF A PCR REACTION, PREPARATION OF SINGLE AND DOUBLE DIGESTIONS FOR THE CONSTRUCTION OF A RESTRICTION MAP. CONTROL OF ENZYMATIC DIGESTIONS AND PCR BY AGAROSE GEL ELECTROPHORESIS (4 HOURS OF LABORATORY EXERCISE FOR GROUPS OF 2/3 PEOPLE) -PREPARATION OF A LIGASE REACTION, TRANSFORMATION INTO COMPETENT CELLS, PLATING ON SELECTIVE SOIL. CONSTRUCTION OF A CALIBRATION LINE WITH LEAST SQUARES METHOD, EXTRAPOLATION OF THE MOLECULAR WEIGHTS OF THE DIGESTION FRAGMENTS (4 HOURS OF LABORATORY EXERCISE FOR GROUPS OF 2/3 PEOPLE) -DISCUSSION OF THE RESTRICTION MAP. CONTROL OF RECOMBINANT COLONIES. PLASMID DNA EXTRACTION BY ALKALINE LYSIS (4 HOURS OF LABORATORY EXERCISE FOR GROUPS OF 2/3 PEOPLE). |
| Teaching Methods | |
|---|---|
| THE COURSE INCLUDES 36 HOURS OF THEORETICAL LECTURES THAT TAKE PLACE IN THE CLASSROOM WITH THE HELP OF POWERPOINT PROJECTIONS AND 1 2 HOURS OF EXERCISES THAT TAKE PLACE IN THE LABORATORY AND INCLUDE THE EXPERIMENTAL DEEPENING OF THE BASIC TECHNIQUES OF MANIPULATION AND ANALYSIS OF NUCLEIC ACIDS. |
| Verification of learning | |
|---|---|
| THE ACHIEVEMENT OF THE OBJECTIVES OF THE COURSE IS CERTIFIED BY PASSING AN EXAM WITH EVALUATION IN THIRTIETHS. THE EXAM INCLUDES AN ORAL TEST, OF ABOUT 30 MINUTES, DURING WHICH RESEARCH PROBLEMS CONCERNING THE PRODUCTION OF BIOTECHNOLOGICAL DRUGS, GENE EXPRESSION VARIATIONS AND THE PRODUCTION OF GENETICALLY MODIFIED ORGANISMS WILL BE PROPOSED; THE STUDENT WILL HAVE TO DEMONSTRATE TO BE ABLE TO FOCUS ON THE PROBLEM AND PROPOSE THE APPROPRIATE RESEARCH STRATEGY FOR ITS RESOLUTION. THE MINIMUM MARK FOR PASSING THE EXAM IS 18/30 AND WILL BE ATTRIBUTED TO THE STUDENT WHO SHOWS KNOWLEDGE OF THE METHODS, BUT LIMITED ABILITY TO FOCUS ON THE SCIENTIFIC PROBLEM AND PROPOSE A RESEARCH STRATEGY. PRAISE CAN BE ATTRIBUTED TO STUDENTS WHO REACH THE MAXIMUM MARK OF 30/30 AND DEMONSTRATE TO HAVE COMPLETE MASTERY OF TECHNOLOGIES AND RESEARCH AND ANALYSIS STRATEGIES IN THE BIOTECHNOLOGICAL FIELD. |
| Texts | |
|---|---|
| BROWN T.A.-MOLECULAR BIOTECHNOLOGIES-ZANICHELLI-ISBN: 978-88-08-16746-0 RECOMMENDED TEXTS FOR FURTHER INFORMATION: REECE R.J.-ANALYSIS OF GENES AND GENOMES -EDISES- ISBN: 8879593463 AMALDI ET AL., TECHNIQUES AND METHODS FOR MOLECULAR BIOLOGY-CEA- ISBN: 978-88-08-92034-8 |
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