Teacher
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GUALANDI Giampiero
(syllabus)
Basic techniques in genetic engineering: - Extraction and purification of chromosomal and plasmid DNA - Properties and use of enzymes which metabolize DNA - Selection of "in vitro" recombinant DNA: chemical and genetic methods - Nucleic acids probes : choice, preparation and labelling - Technique for NA analysis: Southern and Northern Blots, restriction maps and deletions maps - Vectors and hosts for genomic or cDNA libraries - Screening of libraries : DNA/oligo probes, antibodies, subtraction libraries - Cloning by function, complementation, position, tagging. - mapping of structure/function of genomics clones - sub cloning and DNA sequencing - transcripts analysis and mapping - Mapping of regulative regions -"In vitro" mutagenesis - PCR: principle, recent developments and applications - Biotechnological applications
Gene transfer: -Scopes and analysis Cloning and gene transfer in fungi: - Mutants, biological cycles, sexual and parasexual in yeast and aspergillus. Complementation, mapping, recombination (mechanisms) - trasformation , vectors in yeast and functional elements of chromosome (ARS,CEN, TEL) Artificial chromosomes Plasmid 2 micron - Techniques for gene disruption and gene replacements in yeast - Gene cloning and trnasfer in aspergillus Higher eukaryotic systems - Cell cultures, mutants and markers for selection. Techniques for DNA delivering. - Animal viruses, genetic characteristics , derived vectors for reolicating plasmids and for trasduction: SV40, herpesvirus, AAV, retroviruses - Integration of vectors and delivered DNA: illegittimate recombination - Methods for tagging: selections, PCR. Use of site specific recombination. KO technology, CRISPR and RNA interference - Transgenic animals - General features and problem for gene therapy
(reference books)
T.Brown, Biotecnologie molecolari , Zanichelli - S.Kingsman e A.Kingsman, Ingegneria Genetica, Piccin ed. - S.Primerose, R.Twyman,B: Old, Ingegneria Genetica. Zanichelli - Reviews and scientific papers provided by the teacher
Slides available at the link: https://www.dropbox.com/s/z98dr3zcgj0pxlz/Ing%20Gen%20proc%208.pptx?dl=0
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