Topic |
Sub-topic |
1.- GENOME ORGANIZATION |
C-value paradox. Prokaryotic and eukaryotic genomes. Single-copy and repetitive DNA sequences. Gene families. Centromeres. Telomeres. Organelle genomes. |
2.- DNA REPLICATION |
Semiconservative DNA replication: the Meselson and Stahl experiment. Modes of replication. Enzymology of the replication. DNA replication in Escherichia coli. DNA replication in eukaryotes. Telomere synthesis. Replication of mitochondrial and chloroplast DNA.
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3.- SYNTHESIS AND PROCESSING OF RNA |
Classes of RNA. RNA polymerases. Promoters and transcriptional apparatus. Transcription in prokaryotes and eukaryotes: initiation, elongation and termination. Interrupted genes: exons and introns. Processing of eukaryotic pre-mRNA. Synthesis and processing of pre-rRNA. Synthesis and processing of pre-tRNA. RNA edition. Revision of gene concept.
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4.- TRANSLATION |
The one gene-one enzyme hypothesis. The genetic code: characteristics and experiments to decipher the code. Initiation of translation. Elongation of the polypeptide chain. Termination of translation. Messenger RNA surveillance.
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5.- MUTATION AND DNA REPAIR |
Molecular basis of spontaneous mutations: replication errors, unequal crossing over, spontaneous chemical changes. Molecular basis of induced mutations: chemical and physical agents. Repair mechanisms: direct reversal of damaged DNA, excision repair, mismatch repair, repair of double-strand breaks, translesion synthesis.
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6.- MOLECULAR MECHANISM OF GENETIC RECOMBINATION
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The role of genetic recombination. Gene conversion. Models of homologous recombination: Holliday model and double-strand break model. Enzymes required for recombination. Site-specific recombination. Immunoglobulin gene rearrangements.
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7.- TRANSPOSABLE GENETIC ELEMENTS |
Transposable elements in prokaryotes: insertion sequences, composite transposons and noncomposite transposons. Replicative and non replicative transposition. Transposable elements in eukaryotes: transposons and retrotransposon. Evolutionary significance of transposable elements.
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8.- RECOMBINANT DNA TECHNOLOGY |
Restriction enzymes. Cloning vectors. DNA libraries: construction and screening. Southern and northern blotting. PCR. Restriction maps. DNA sequencing. Site-directed mutagenesis.
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9.- APPLICATIONS OF RECOMBINANT DNA TECHNOLOGY
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Expression of eukaryotic genes in E. coli. DNA transfer to eukaryotic cells. Transgenic animals. Transgenic plants. Gene therapy. Molecular markers. DNA fingerprinting. Genetic diagnosis. Synthetic genomes. Genome editing: CRISPR/Cas9 technology.
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10.- GENOMICS
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Physical and genetic mapping. Whole genome sequencing. Genome annotation. DNA microarrays. Reverse Genetics. Comparative genomics. Metagenomics.
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11.- REGULATION OF GENE EXPRESSION IN BACTERIA
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Jacob and Monod’s operon model for the regulation of lac genes in E. coli. Positive control of the lac operon. The arabinose operon of E. coli: positive and negative control. The triptophan operon of E. coli: negative control and attenuation. Control by RNA molecules. |
12.- REGULATION OF GENE EXPRESSION IN EUKARYOTES
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Changes in chromatin structure. DNA methylation. Transcriptional control.
RNA processing control. Control of mRNA stability. Control at the level of translation. RNA interference. Epigenetics.
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13.- GENETIC CONTROL OF DEVELOPMENT
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Basic events of development. Drosophila development stages. Maternal-effect, segmentation and homeotic genes in Drososphila. Homeobox genes in other organisms. General aspects of Caenorhabditis elegans development. Genetic control of flower development in Arabidopsis. |
PRACTICE 1: DNA EXTRACTION |
DNA extraction from Drosophila melanogaster and human cells. Agarose gel electrophoresis of DNA. DNA quantification. |
PRACTICE 2: PCR |
PCR amplification of the locus PV92. Analysis of an insertion polymorphism of Alu sequences |
PRACTICE 3: DOT-BLOT |
Nucleic acids hybridization: detection of microsatellite sequences by dot-blot |
PRACTICE 4: BIOINFORMATICS. |
Database search and comparison of nucleic acid sequences. Primer design. Identification of ORFs. |