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.
(iv) to be able to read scientific papers developing a critical attitude; and
(v) to develop of a good performance in oral communication of scientific matters in a clear and synthetic form.
Isabel Maria Godinho de Sá Nogueira
Weekly - 4
Total - 63
Mechanisms of genetic changes: mutation; recombination; gene transfer mechanisms; bacterial and phage genetic analysis.
Recombinant DNA Technology - restriction enzymes and restriction mapping; cloning vectors; enzymes used in molecular cloning; 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 characterization 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: Mutation reversions in Bacillus subtilis mutants after different mutagenic treatments. Plasmids transfer by conjugation between of E. coli strains. Gene expression systems. Restriction mappings; cloning strategies; databases to search sequences and use of bioinformatics tools for sequence analysis and comparison; identification of promoter regions, prediction of operons; cellular localization of proteins; identification of point mutations; identification of transposition mutants; construction of transcriptional fusions and fusions of promoters in silico; cloning strategy for construction of conditional mutants.