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MOL 2022: Molecular biology: gene technology and its application
Topics in this unit include recombinant DNA technology; use of bioinformatic tools to handle biological data generated by DNA and protein sequencing; genetic technology and its application in biotechnology. Lecturing Staff
Organisation of the unitMOL2022 consists of 3 lectures per week and one 3 hour practical session per week. The unit is a collaborative course run across three departments: Microbiology, Biochemistry and Molecular Biology, and Biological Sciences. Students will be exposed to laboratory methods in each of the three departments, and classes will include laboratory exercises, computer exercises tutorials, self-directed exercises, oral presentations and written reports. Topics coveredTheme A: Recombinant DNA TechnologyINTRODUCTION TO MOL2022 AND GENE MANIPULATIONBasic cloning requirements. Cutting and joining DNA fragments; vectors; transformation and selection. CLONING STRATEGIES, LIBRARY CONSTRUCTION AND SCREENINGHybridisation. DNA denaturation, hybridisation: liquid and solid phase; labelling of the probe; detection systems; Southern blotting; Northern blotting. CHEMICAL SYNTHESIS, AMPLIFICATION, SEQUENCING AND MUTAGENESIS OF DNAChemical synthesis of DNA. Uses of synthesised oligonucleotides. EXPRESSION OF CLONED GENESProkaryotic expression systems. Manipulation of cloned genes to achieve expression of proteins. Theme B: Bioinformatics – Accessing and Interpreting Gene SequencesBIOLOGICAL DATABASESOverview of the genome project. ANNOTATION OF GENOMIC SEQUENCESHow computers can be used to interpret large sequences. Searching for open reading frames (ORFs). Discussion of genes based on codon bias and size of ORF. Searching databases with derived sequences from ORFs. SEQUENCE ALIGNMENTSSequence alignments. Similarity vs identity. Structural homology in proteins. Construction of multiple alignments and their interpretation. Structural implications of amino acid conservation.Using sequences to search the databases : BLAST. PHYLOGENYEvolution and its relationship to conserved residues. Conservative and non-conservative substitutions. PROTEIN STRUCTURE AND MODELINGFrom protein sequence to structural information. Homology modelling of protein structure and comparison with experimental methods. Theme C: Applications of Recombinant DNA -Molecular BiotechnologyApproaches for genetic manipulation of microbes, plants and animals.Manipulation of DNA sequence to alter gene structure (PCR based mutation) Concepts of totipotency in eukaryotes (Fungi, Plants, Animals/vertebrates) Introduction of foreign DNA into genomes by physical methods (microinjection, biolistics, electroporation, liposomes) Introduction of foreign DNA into genomes of fungi, plants and animals using specialized vectors (bacterial, viral) Copy number determination and inheritance of foreign genesGene targeting vs random insertion of foreign DNA sequences Strategies for down regulation of gene expression (antisense genes, oligonucleotides, gene knockouts) RNA interference via double stranded RNA and targeted mutation using TILLING Studying expression of foreign genes using reporter genes (GUS, GFP, CAT, Luc) Examples of uses of transgenic microbes, fungi and animals.Therapeutic proteins, vaccines and antibodies Manipulation and synthesis of antibodies, small biologically active molecules and novel industrial bioproducts such as biopolymers and oils/solvents Bioremediation, biotransformation and degradation of xenobiotics Molecular breeding to improve pest/disease resistance and sustainable production by organisms/species of agricultural and industrial importance (yeast; fish; crop plants; animals) Gene therapy – applications in medicine (cystic fibrosis; ADA deficiency; possibilities for treatment of cancer) Release of transgenic organisms, products and patenting/regulatory issues arising from this technology Ethical and safety issues for the environment and consumers (“GMO debate”) |