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MOLECULAR BIOLOGY (A-L)
ANNA LA TEANA

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

Prerequisites

Knowledge of basic General Chemistry, Cytology, Organic Chemistry and Biological Chemistry is required.

Development of the course

The course is organized in lectures (7 CFU, 56 hours) and laboratories (1 CFU, 8 hours). Handouts of the lectures, protocols for the laboratories and booking forms for the laboratories are loaded on the Moodle platform of DiSVA website.

Learning outcomes

Knowledge:
Students will learn about the structure-function relationship of nucleic acids and of nucleic acids interacting proteins, the molecular mechanisms of DNA duplication, RNA transcription and maturation, translation and regulation of gene expression. The course will also provide a description of the different experimental procedures, which have led to current knowledge.

Ability to apply the knowledge:
Students will be able to apply the acquired knowledge about the working principles of genetic material to other topics such as Genetics, Developmental Biology and Cell Biology. Furthermore, they will be able to apply some basic DNA manipulation and analysis techniques; they will be able to search nucleic acid and protein sequences in databases and to use software specific for restriction analysis and site-directed mutagenesis of DNA fragments.

Soft skills:
The practical experience during the laboratories together with the discussions on the experimental results will improve students’ autonomy and ability to evaluate critically scientific data. Furthermore, the organization of the laboratories in small groups will facilitate the coordination and communication abilities.

Program

Content (lectures 7 CFU, 56 hours):
Nucleic acids. Structure and chemical-physical properties. Nucleic acids as genetic material. DNA topology. Structural organization of viral, prokaryotic and eukaryotic genomes. Chromosomes, chromatin, nucleosomes.
DNA replication. The Meselson and Sthal experiment. The replication fork and the semidiscontinuous synthesis of DNA. Coordinated synthesis of the leader and lagging strands. DNA polymerases in prokaryotes and eukaryotes. Replication origins. Regulation of replication initiation in prokaryotes and eukaryotes. Replication and cell cycle.
DNA recombination. Homologous and site-specific recombination. Transposition.
Gene organization in virus, prokaryotes and eukaryotes.
Transcription. Different types of RNA: mRNA, tRNA, rRNA, snRNA, scRNA.
Transcription of prokaryotic genes. RNA polymerase and promoters. Termination and anti-termination.
Transcription of eukaryotic genes. RNA polymerases and promoters. Transcription factors. Enhancers and silencers. Termination.
RNA processing. Processing of rRNA and tRNA. mRNA maturation and splicing. Self-splicing. Editing.
mRNA translation. tRNA as an adaptor: secondary and tertiary structure. Modified bases. The genetic code. The aminoacyl-tRNA synthetases and the identity rules.
The ribosome. The different steps of protein synthesis. Initiation, elongation and termination factors in prokaryotes and eukaryotes. The role of rRNA in protein synthesis. Antibiotic and protein synthesis.
Regulation of gene expression in prokaryotes. The operon. Structural genes and regulator genes. Induction and repression: the lac, trp, examples. Catabolite repression. Attenuation. Examples of regulation at the post-transcriptional level.
Regulation of gene expression in eukaryotes. Response elements and DNA binding protein domains. Different models for gene activation. DNA methylation and gene expression. Chromatin structure and transcription.
Experimental procedures
DNA analysis: digestion with restriction enzymes, restriction mapping, cloning vectors, DNA sequencing, PCR, Southern blotting, site-directed mutagenesis. Promoters analysis: footprinting and band-shift, reporter genes, mutations analysis. Transcripts analysis: northern blotting, 5’-race. mRNA purification by oligo-dT and cDNA libraries construction. Translation analysis: Cell-free systems, Western blot.

Laboratories (1 CFU, 8 hours):
Students will perform: extraction of plasmid DNA from bacteria, purification of the plasmid by affinity chromatography and determination of its concentration by spectrophotometric analysis; restriction enzymes digestion and analysis of the fragments by agarose gel electrophoresis; data analysis. Search for nucleic acid and protein sequences in the NCBI database, identification of a gene coding sequence, in silico restriction mapping and site-directed mutagenesis, sequence alignment using BLAST. 

Development of the examination

Methods for assessing learning outcomes:
Evaluation is performed through an oral exam during which students will be asked to discuss about 3 different topics.

Criteria for assessing learning outcomes:
Questions aim at verifying the level of knowledge acquired by the students. They span the entire program: topics discussed during the lectures as well as topics subject of the laboratories.

Criteria for measuring learning outcomes:
The level of knowledge acquired by the students is measured with a mark between 0 and 30. In order to pass the exam the final mark must be between 18 and 30. The highest possible mark is 30/30 cum laude.  

Criteria for conferring final mark:
The answer to each question is evaluated with a mark included between 0 and 10. The final mark is calculated as the sum of the 3 answers. 30/30 cum laude is attributed to students particularly able to discuss critically and with great competence about the different topics.

Recommended reading

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

Courses

• Scienze biologiche