It is intended that at the end of this course students understand in what consist strategies that are based on recombinant DNA technology, which are the tools used in gene manipulation; how this technology encompasses multiple domains of life sciences, and some of its current applications. It is intended that the students gain knowledge of some of the recombination technologies which allow gene cloning, construction of genomic libraries, mapping and identification of genes, gene expression analysis, the expression of heterologous genes and directed mutation.
DNA recombination in vitro: restriction enzymes and ligases; cloning vectors and cloning of genes.
Methods for introducing recombinant DNA into cells and selection of clones.
Construction of genomic libraries.
Homologous and heterologous gene expression: Systems / expression vectors.
Analysis of the level of gene expression.
Detection and subcellular localization of proteins encoded by cloned genes: immunodetection and GFP-fusions.
Directed mutagenesis and protein engineering. Interruption and deletion of genes.
RNAi technology.
Applications in Healthcare, Biotechnology, Agriculture and Environment.
The TP component integrates the presentation of the theoretical foundations included in the program. A methodology for interactive exposition, encouraging discussion of the topics presented is followed. Examples and problematic issues for discussion and resolution by the students are presented, using bioinformatic tools and databases.
In OT classes is intended that students discuss some practical protocols that allow the cloning and expression of genes in Escherichia coli. It is also intended that students perform some tasks of those protocols, in order to consolidate the knowledge acquired during the TP.
T.A. Brown, 2010. Gene cloning and DNA analysis : an introduction. 6ª ed. Wiley-Blackwell.
Sambrook & Russel. 2001. Molecular Cloning: A Laboratory Manual. 3ª ed. Cold Spring Harbor Laboratory Press.
S.B. Primrose & R.M. Twyman. 2006. Principles of gene manipulation and genomics. 7ª ed. Blackwell Publishing.
Desmond S. T. Nicholl. 2008. An Introduction to Genetic Engineering. 3ª ed. Cambridge University Press.
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