PHA.06110.02 - C-Both: Construction of production organisms - Hosts and vectors (Vollständige Modulbeschreibung)

S. Schilling

Course C-Both.1: Methods and strategies for the generation of production strains

• Overview biotechnologically relevant organisms: product related selection, advantages/disadvantages
• Toolbox biotechnology: Gene analysis, DNA/protein sequencing, detection methods for products
• Strategies for improvement of strains and products: random/site directed mutagenesis, protein tags
• Examples for successful application in pharmaceutical and industrial biotechnology using different production organisms (bacteria, yeast, plants)

Course C-Both.2: Biotechnology of phototrophic organisms

• Principal organization and expression of nuclear and organellar plant genomes
• Plant and cell architecture, compartments and protein sorting
• Strategies and procedures for plant regeneration and propagation
• Plant cell, tissue and organ cultures: initiation and maintenance
• Mechanism of Agrobacterium-mediated T-DNA transfer and applications
• Plant transformation techniques and gene expression systems in plants
• Vector design and optimization, stability and analysis of transgenes
• Examples of molecular plant biotechnology: natural and novel products
• Overview of (biotechnological relevant) micro- and macroalgae and application
• The unicellular green algae Chlamydomonas reinhardtii as a model organism
• Molecular biology of and gene technology with Cyanobacteria

Course C-Both.3: Molecular Biotechnology

• Principal mechanisms of DNA synthesis in vitro and in vivo and use of enzymes for gene cloning
• Mechanism of RNA and protein synthesis in bacteria
• Structure and function of RNA and DNA
• Regulation of gene expression in prokaryotes and eukaryotes (yeast) and use for heterologous expression of proteins
• Methods of gene cloning and target gene isolation, library construction and screening
• Theoretical ability to construct a genetically modified organism for heterologous protein production
• Theoretical ability to cultivate bacteria and yeasts

Course C-Both.4: Project seminar 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.
• Testing transgenic plants for the presence of transferred DNA
• Monitoring heterologous protein production in plants

Examination on course contents part microorganisms, Protocol on course part microorganisms, Examination on course contents part plant cell technology, Protocol on course part plant cell technology

Klausur