12946 - Molecular Biology

Learning outcomes

After completing this course, the student has adequate knowledge of the fundamental principles and basic mechanisms of molecular biology, and of the fundamental nuclear mechanisms of gene expression in the eukaryotic and prokaryotic cells. In particular, the student is able to understand and discuss issues of gene expression, replication and DNA repair, and the structure of the gene at the molecular level. Moreover, the student knows basic mechanisms operating in the eukaryotic cell, including transcription and its regulation, transcript maturation and its regulation, the structure and regulation of chromatin, gene organization and general principles of genome organization. At the end of the laboratory the student knows how to perform basic molecular biology techniques, such as PCR, DNA restriction and ligation, construction of recombinant plasmids, Escherichia coli transformation and heterologous protein expression.

Course contents

Module 1 (5 cfu, 44 hr)

Structure, components and properties of nucleic acids. Gene structures. Central dogma. Genetic code. tRNA structure and synthetases. Ribosome. Protein synthesis: stages of translation initiation factors in prokaryotes and eukaryotes, elongation,translocation, termination. Codon-anticodon interactions. Transcription in prokaryotes: The RNA polymerase and transcription. Structure and function of sigma factors. Structure of the promoter. Termination. Antitermination. Bacterial operons andcoordinated regulation: lac, trp. Structure and function of the Lacrepressor. Operators. Catabolite repression. Antisense RNA. Phage strategies. Phage lambda. Structure of the CI repressor and lysogeny. Comparison of lytic and lysogenic cycles. Transcription in eukaryotes: RNA polymerase, transcription factors and regulatory elements. Replication. DNA polymerase I and III of E. coli. Other DNA polymerases. Repair systems. Recombination. Rec and Ruv systems. DNA topology.

 

Module 2 (40 hr)

Gene organization. Introns and exons, pseudogenes, gene clusters (globin genes and ribosomal genes). Structure and function of repeted DNA sequences. Transcription. Classes of RNA polymerases. Structure of RNA polymerase II. Promoter organization of class I, II and III. General transcription factors and their funztion. TBP structure in the DNA complex. Transcription regulation. Inducile transcription factors. Enhancers. Protein intaraction domains with DNA: structure and function. Chromatin. Chromatin structure. Nucleosome: X-ray structure and molecular components. Post-translational modification of histones. Translational and rotational nucleosomal positioning. Chromatin role in transcription regulation. Transcript maturation. Intron classes. Splicing mechanism. Autosplicing mechanism of group I and II introns. Molecular mechanism of tRNA splicing.

 

Module 3 (laboratory, 2 cfu, 30 hr)

Laboratory practical course module

PCR goals and applications: amplification and modification of a DNA template.

DNA and protein electrophoresis

Generation of recombinant plasmids: restriction and ligation reactions.

Expression of eukaryotic recombinant proteins in a heterologous prokaryotic system: the T7 system.

Readings/Bibliography

Module 1: Il Gene, Seconda Edizione Compatta, B. LEWIN, Zanichelli. Chapters: 1-3, 7-9, 11-15, 18-20

The slides presented in class do not replace the reference text but is a guide for the selection and understanding of the topics to be studied.

Module 2: Capranico G, Martegani E, Musci G, Raugei G, Russo T, Zambrano N, Zappavigna V. Biologia molecolare, EdiSES (ultima edizione)

Teaching methods

Class lectures

The course consists of three modules (10 CFU) of lectures aimed to the comprehension of the fundamental principles and basic mechanisms of cell functioning.

The laboratory course consists of 2 CFU, corresponding to 30 hours of practical work. Each student will performs the experimental protocol individually, thought working in groups of 4-5 students. During the course, exercises will be be given to students in order to strengthen the theory underneath lab techniques.

As concerns the teaching methods of this course unit, all students (including all the international incoming exchange students, i.e. ERASMUS) must attend Module 1, 2 online [https://www.unibo.it/it/servizi-e-opportunita/salute-e-assistenza/salute-e-sicurezza/sicurezza-e-salute-nei-luoghi-di-studio-e-tirocinio], while Module 3 on health and safety is to be attended in class. Information about Module 3 attendance schedule is available on the website of your degree programme ("studiare"--"formazione obbligatoria su sicurezza e salute").

Assessment methods

Module1: Written test

The exam takes place only through the final exam, which ensures the acquisition of knowledge and expected skills by conducting a written test for 1.5 hours without the aid of notes or books.

The written test consists of 40 multiple-choice questions and one open question. Each multiple-choice question achieves 0.5 points, while the open question allows for a maximum of 10 points. The final grade of the module is calculated as the sum of the obtained points.

Module 2: Written test

The evaluation is out of thirty. Registration on Alma Esami is required for this module. The exam consists of a written test. The written test consists of multiple-choice quizzes and open-ended questions. The total of the scores is greater than 30 and therefore allows to obtain the praise. The test is passed if the total score is equal to or greater than 18.

Laboratory practical course module: the verification test consists of a written report of the experience carried out in the laboratory. The student will have to write a summary report in which the different passages of the experience are reported and their importance for the final result is explained. The report must be delivered by the date established during the laboratory activities. The results of the exercises delivered and carried out during the module and the student's behavior in the laboratory are also taken into consideration in the final grade. 

 

The final mark of the teaching will be the weighted average of the marks obtained in the three modules.

Teaching tools

PC, video projection and board.

In the lab all the instruments and reagents necessary for the experimental procedures are provided to students.

Office hours

See the website of Vincenzo Scarlato

See the website of Jessica Marinello

See the website of Jessica Marinello