PHA.03533.03 - Construction of production organisms: Hosts and vectors (Vollständige Modulbeschreibung)

PD Dr. M. Brandsch

• To understand the basic principles of cell physiology. • To obtain solid knowledge on morphology and function of cell organelles. • To understand the basic techniques of in vitro animal cell cultures. • To know the problematic nature of stem cell research. • To obtain detailed knowledge on transfection of mammalian cells for biotechnological purposes. • To understand the basic principles of plant development • Knowledge of the basic concepts of in vitro plant cell, tissue and organ cultures • Insight into molecular techniques of plant cell transformation and regeneration • Overview of industrial application of plant tissue systems • Insight into biotechnological applications of transgenic plants • Survey of basic techniques of in vitro cultivation of plant cells • Skills of induction and cultivation of plant cells • Skills of handling agrobacteria in plant transformation procedures • Practical knowhow in analyzing genetically modified plants • Theoretical ability to construct a genetically modified organism by gene cloning und ex-pression in a final production host. • Advantages of bacteria, yeast and insect cells as gene expression systems. • Theoretical ability to cultivate bacteria and yeasts. • Overview over products of molecular and classic biotechnology. • Practical know-how for transformation/transfection and cultivation of plant and microbial cells

Course C.1: Animal cell biology and technology • Animal cell morphology and biochemistry • Animal cell physiology • Animal cell lines: Adherent cells, cells in suspension • Basic cell culture techniques: Trypsinization, subculturing, proliferation and viability tests • Transfection techniques • Animal cell mass culture • Special cell types: hybridoma cells
Course C.2: Plant cell technology • Plant meristems, vegetative growth and reproductive development • Basic plant regeneration and propagation procedures • Application of organ culture systems • In vitro storage and cryopreservation of plant tissues • Plant cell transformation techniques • Gene expression systems in plants • Molecular plant biotechnology: natural and novel products • Plant cell, tissue and organ cultures: initiation and maintenance • Monitoring of growth parameters and proliferation of plant cell cultures • Agrobacterium-mediated transient expression of reporter genes • Testing transgenic plants for the presence of foreign DNA by PCR • Monitoring of promoter activation based on reporter enzyme activity
Course C.3: Molecular Biotechnology • Principal mechanisms of DNA synthesis in vitro and in vivo - use of enzymes for gene cloning. • Regulation of gene expression in prokaryotes and use for heterologous expression of proteins. • Methods of gene cloning and target gene isolation. • Theoretical ability to construct a genetically modified organism for heterologous protein expression. • Theoretical ability to cultivate bacteria and yeasts. • Advantages of bacteria and yeast as gene expression systems.
Course C.4: Practical course on construction of hosts and vectors • Amplification of target DNA using PCR, restriction digest and vector ligation. • Transformation of E. coli with plasmid, selection, monitoring of the expression success.

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Klausur