Dipartimento di Scienze della Vita e dell'Ambiente - Guida degli insegnamenti (Syllabus)

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MOLECULAR BIOLOGY (M-Z)
DAVIDE SARTINI

Seat Scienze
A.A. 2016/2017
Credits 8
Hours 64
Period 2^ semestre
Language ENG
U-gov code ST01 3S038

Prerequisites

Students are supposed to have acquired principles of General and Inorganic Chemistry, Cytology, Organic Chemistry and Biochemistry.



Development of the course

The course will include frontal lectures (7 CFU, 56 hours) and practical activities in experimental and informatic laboratories (1 CFU, 8 hours). Within DiSVA website, the Moodle platform will supply students with teaching materials of frontal lectures, experimental and informatic laboratory protocols and reservation forms to participate to practical activities.



Learning outcomes

Knowledge:
The course topics, covered during frontal lessons as well practical activities, will allow students to learn concepts about structure and function of nucleic acids, protein–DNA and protein-RNA interactions, molecular mechanisms underlying DNA replication, transcription, translation and RNA processing, and regulation of gene expression.

Ability to apply the knowledge:
Student will apply knowledges, acquired during the course, to learn other subjects such as Genetics and Developmental Biology. Student will also be able to perform basic procedures used for manipulation and analysis of nucleic acids, to find specific nucleic acid and protein sequences stored in databases available through web and to use specific softwares for restriction analysis and site-directed mutagenesis.



Program

Frontal lectures (7 CFU, 56 hours).
Nucleic acids. Nucleic acids as genetic material. Structure and physical and chemical properties of DNA and RNA. DNA topology. Organization of viral, prokaryotic and eukaryotic genomes. Chromosomes, chromatin and nucleosomes.
DNA replication. The Meselson and Stahl experiment. The replication fork. The semi-discontinuous DNA replication The coordinated synthesis of both leading and lagging DNA strands. Prokaryotic and eukaryotic DNA polymerase. Replication origins. Regulation of DNA replication initiation in prokaryotes and eukaryotes. Replication and cell cycle.
Transcription. Different types of RNA molecules: mRNA, tRNA, rRNA, and snRNA. Transcription in prokaryotes. Transcription initiation: RNA polymerase and promoters. Intrinsic and Rho-dependent termination. Anti-termination. Transcription in eukaryotes. Transcription initiation: promoters and consensus sequences. RNA polymerase I, II and III. Transcription factors. Enhancers and silencers. Transcription termination. Processing of rRNA, tRNA and mRNA. Splicing: spliceosome, snRNA and snRNP. Self-splicing: group I and II introns. RNA editing.
Translation. tRNA as an adaptor molecule: secondary and tertiary structure. The genetic code. Aminoacyl-tRNA synthetase. The ribosome. Steps in protein synthesis. Initiation, elongation and termination factors in prokaryotes and eukaryotes. The role of rRNA in protein synthesis. Antibiotics and protein synthesis.
Regulation of gene expression in prokaryotes. The operon. Structural and regulatory genes. Induction and repression: lac and trp operons. Catabolite repression and attenuation. Regulation of gene expression in bacteriophage λ.
Regulation of gene expression in eukaryotes. Response elements. DNA binding domains. DNA methylation and gene expression. Chromatin structure and transcriptional activity.
Methods. Molecular cloning: restriction enzymes and cloning vectors. PCR and agarose gel electrophoresis. DNA sequencing. Blotting methods: Southern blot, Northern blot and Western blot. Site-directed mutagenesis. Purification of mRNA by oligo(dT)-cellulose chromatography. Genomic and cDNA libraries. Expression vectors and recombinant proteins.

Practical activities in experimental and informatic laboratories (1 CFU, 8 hours).
Isolation of plasmid DNA from bacterial cells. Evaluation of plasmid DNA concentration through spectrophotometric analysis. Digestion of plasmid DNA using restriction endonucleases. Separation of DNA fragments by agarose gel electrophoresis.
Use of nucleotide and aminoacid sequence databases availabe at NCBI. Identification of the coding sequence of a structural gene. Restriction map and site-directed mutagenesis through in silico analyses. BLAST as a tool for sequence allignment.



Development of the examination

Methods for assessing learning outcomes:
Learning assessment will be performed through an oral exam. During oral exam (30 minutes) each student will be asked 3 questions about topics within the program content.

Methods for assessing learning outcomes:
Oral exam questions will evaluate the knowledge of topics dealt with during frontal lectures as well as practical activities in experimental and informatic laboratories.

Criteria for measuring learning outcomes:
The final mark is expressed as a ratio. The exam is passed when the ratio is equal o more than 18/30. It is possible to attribute full mark with honour (30/30 cum laude).

Criteria for measuring learning outcomes:
Each answer will be assigned a score ranging from 0 to 10. The final result of the oral exam is calculated as sum of the scores obtained for each answer. Full mark with honour could be attributed if the student demonstrate mastery of the body of knowledge and high level communication skill.



Recommended reading

Biologia Molecolare, F. Amaldi, P. Benedetti, G. Pesole, P. Plevani. Casa Editrice Ambrosiana. II ediction. 2014.
Biologia molecolare del gene, J.D. Watson, T.A. Baker, S.P. Bell, A. Gann, M. Levine, R. Losick. Casa Editrice Zanichelli. VII ediction. 2015.



Courses
  • Scienze biologiche




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