Molecular Biology B
Code
1116
Academic unit
Faculdade de Ciências e Tecnologia
Department
Departamento de Ciências da Vida
Credits
6.0
Teacher in charge
José Paulo Nunes de Sousa Sampaio, Sérgio Joaquim Raposo Filipe
Weekly hours
4
Total hours
72
Teaching language
Português
Objectives
The objective of this discipline is the integration of basic knowledge about the synthesis, structure and function of nucleic acids and regulation of gene expression. Also, to encourage awareness of the different applications of the molecular biology and interdisciplinarity with other sciences. To familiarize students with basic concepts and fundamentals of methods of nucleic acids manipulation and analysis and to use databases, such as of nucleic acid or protein sequences.
Prerequisites
Laboratory/practical lectures are mandatory with participation in problem solving sessions.
Subject matter
Lectures: Topic 1. DNA-Chromosomes-Genome: Concepts of structure and function of DNA, chromosomes and genomes. Topic 2. DNA Replication, and Repair : Chemistry of DNA synthesis. Replication enzymes (DNA polymerases, transcriptases, reverse transcriptases). Mechanisms of replication of circular and linear DNA molecules. Topic 3. From DNA to protein: Fundamentals of the mechanisms of transcription and translation.Topic 4. The concept of spontaneous versus induced mutation and brief introduction with examples of DNA repair systems. Topic 5: Regulation of gene expression in prokaryotes (E. coli model) at transcription level: lactose operon, the tryptophan operon and plambda phage regulation. The role of small non-coding RNAs in gene expression in bacteria and eukaryotic organisms.
Laboratory classes: Growth and culturing of bacteria in liquid media. Preparation of serial dilutions for count colonies and determination of colony-forming units. Isolation, purification, and quantification of nucleic acids: plasmid DNA by alkaline lysis. UV spectrophotometry. Enzymatic hydrolysis of plasmid DNA. Agarose gel ectrophoresis to visualize isomeric forms of circular DNA molecules and DNA integrity. Physical mapping. PCR. Islation of cheek cell DNA extraction (adapted ‘DNA Extraction Module Genes in a Bottle” lab kit. Bio-Rad Lab.). UV spectrophotometry. Agarose gel ectrophoresis.
Bibliography
- Alberts B., Johnson A., Lewis J., Raff M., Roberts K., Walter P. (2008). Molecular Biology of the Cell (5th ed.). Garland Science.
- Research papers.
Teaching method
Lectures, laboratory and theorical-pratical sessions with quizzes.
Evaluation method
The theoretical and practical-theoretical contents are evaluated by two written tests (75% of final grade) performed individually, 2 oral presentations (20% of final grade), continuous evaluation at the laboratory (5% of final grade) .