Genetic Engineering

Course Description

COURSE CONTENT

• Introduction to Genetic Engineering - Basic concepts in genetic engineering, application scope and implications for human society; comparison of classic breeding methodology and methods of genetic engineering; specific examples of expression of human proteins in E. coli
• Restriction enzymes and recombination "in vitro"
• nucleic acid electrophoresis
• Enzymes for modification of nucleic acids (ligases, polymerases, nucleases, kinases, phosphatases, transferases ...)
• Methods of PCR, qPCR, RAPD and targeted mutagenesis in vitro
• Vectors and hosts in genetic engineering and the construction and search of a gene bank
• Labelling of nucleic acids and application of hybridization methods
• Methods for detection and analysis of DNA polymorphisms
• DNA sequencing and postgenomic research
• Genetic engineering of S. cerevisiae yeast
• Genetic modifications of plants and animals

LEARNING OUTCOMES

• explain the importance and application of genetic engineering in biotechnology and support the explanations by concrete examples
• explain basic concepts related to genetic engineering such as genetic modification, recombinant DNA, cloning, GMOs, genes, transgenes, cDNAs, ORFs, clones, cloning, vector, inserts, transformations, homologous / heterologous expression, libraries / gene bank ...
• apply enzymes for cleavage and modification of nucleic acids such as restriction enzymes, DNA ligases, DNA and RNA polymerases, RNase and DNase, and phosphatase and kinase, for the purpose of constructing and analysing recombinant plasmids, construction of gene bank and genetic material analysis
• explain the principle, procedure and application of the following methods: DNA electrophoresis, restriction analysis of DNA, isolation of DNA from gel, PCR, qPCR (using "TaqMan probes"), RAPD, RFLP, VNTR, AFLP, SSCP, DGGE, FISH, DNA sequencing (dideoxy), S1 mapping, Southern blotting, Northern blotting, two hybrid system, plasmid isolation in singlestranded form and targeted mutagenesis in vitro (by Kunkel method and PCR)
• explain the principle and procedure of cloning and construction of a genomic bank in some vectors such as plasmids, viral vectors, phagmids, cosmids, BACs, PACs and YACs
• plan and propose strategies for the introduction of targeted genetic modification in yeast S. cerevisiae
• explain the principle and procedure for the genetic modification of plants and genetic modification and cloning of animals
• plan and carry out the construction of the plasmid and the gene bank, carry out restriction analysis of plasmid or DNA fragment and perform methods such as PCR, RAPD and RFLP
• interpret the results of molecular genetic analysis

To enrol in this course, the following courses must be completed:

• Molecular Genetics
• Biochemistry 1