Faculdade de Ciências e Tecnologia
Departamento de Ciências da Vida
Teacher in charge
Ana Madalena Peres de Drumond Ludovice Mendes Gomes
The students should be able (i) to understand the natural mechanisms of genetic exchanges at molecular level and their important role on the recombinant DNA technology establishment; (ii) to learn laboratory techniques and methodologies used in genetic engineering; (iii) to integrate the theoretical concepts and the practical approaches, and to design experimental strategies to developed different scientific proposals.
is recommended that the students have a basic background on molecular biology, biochemistry and genetics.
First part - Mechanisms of genetic changes - Mutation; Recombination; Gene transfer mechanisms; 4. Bacterial and phage genetic analysis.Second part: - Recombinant DNA Technology - Restriction enzymes: restriction mapping; molecular typing; forensic genetic; Vectors: plasmids (E.coli and yeast cloning plasmids; shuttle vectors; Ti plasmid of Agrobacterium; expression vectors); viral genomes (l phage; M13 and f1; SV40; retrovirus); cosmids. Enzymes used in molecular cloning; Molecular methods: DNA fragments PCR amplification; probes preparation (genomic, cDNA and oligo probes); labelling (radioactive and non-radioactive methods; prim); molecular hybridisation; in situ hybridisation. Transcription start point determination by nuclease S1 and primer extension. DNA / protein interactions: gel retardation and foot printing. Molecular cloning strategies: genomic and cDNA libraries; shotgun cloning and gene screening; chromosome walking; gene inactivation by RNA anti-sense; in vitro mutagenesis. Production and characterisation of recombination proteins: Lac-fusions and tags-fusions; antibody production; western-blotting. Genomic and proteomic: genetic functional analyses in post-genomic era (transcriptional profiles – arrays; 2D gel electrophoresis).
Laboratory sessions:(1) Detection of Bacillus subtilis mutants after different mutagenic treatments.(2) Plasmids transfer by conjugation between of E. coli strains.(3) gene expression system.
Teaching strategy involves lectures, lab and computer/bioinformatic sessions.Lecture handouts and other learning support are available in the course Moodle webpage.