| Topic |
Sub-topic |
| 1.- 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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| 2.- 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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| 3.- TRANSLATION |
Central dogma in molecular biology. Ribosomes and tRNAs. Translation cycle: initiation, elongation, and termination. Genetic code and genetic decoding. Peptydil transferase reaction. The ribosome: composition. Phylogenetic conservation of rRNA. Role of rRNA in initiation. Role of RNA in decoding. Role of RNA in peptydil transfer. The hypothesis of the RNA world. |
| 4.- 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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5.- 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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| 6.- 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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| 7.- 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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8.- 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. Genetic diagnosis. Genome editing: CRISPR/Cas9 technology.
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9.- GENOMICS
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Structural genomics: molecular markers and genetic maps. DNA fingerprinting. Structural genomics: physical maps and genome annotation. Functional genomics: DNA microarrays, RNA-seq and reverse genetics. Comparative genomics. Metagenomics. Synthetic biology.
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10.- 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. RNA-mediated regulation. |
11.- 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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| PRACTICE 1: DNA EXTRACTION |
Genomic DNA extraction. Agarose gel electrophoresis for DNA. DNA quantification. |
| PRACTICE 2: PCR |
PCR amplification of the CHD gene. Analysis of an intron polymorphism for bird sexing. |
| PRACTICE 3: DOT-BLOT |
Nucleic acids hybridization: detection of microsatellite sequences by dot-blot |
| PRACTICE 4: BIOINFORMATICS. |
Analyses and comparison of nucleic acid sequences. Primer design. |
Subjects