| Department of Plant Biology
80 % Department of Basic Animal and Veterinary Sciences 15 % Department of Food Science 5 % | |||||||||||||||||
| Earliest Possible Year | MSc. 1 year to MSc. 2 year | ||||||||||||||||
| Duration | One block | ||||||||||||||||
| Credits | 15 (ECTS) | ||||||||||||||||
| Course Level | MSc | ||||||||||||||||
| Examination | Final Examination oral examination All aids allowed Description of Examination: The exam will be based on the topics covered in the practical exercises as well as on related theoretical cases. The exact cases will be announced one week before the examination. Weight: The final examination count 100% 13-point scale, internal examiner | ||||||||||||||||
| Requirement For Attending Exam | 75 % of the report for the practical part of the course must be approved in order to attend the final examen | ||||||||||||||||
| Organisation of Teaching | The course contains both a theoretical part and a practical part. In the theoretical part there will be lectures as well as student presentations based on cases and journals. A laboratory course is running several days during most weeks | ||||||||||||||||
| Block Placement | Block 3 Week Structure: Outside schedule | ||||||||||||||||
| Teaching Language | English | ||||||||||||||||
| Optional Prerequisites | 230003 Biochemistry 2 240042 Thematic Course: Experimental Molecular Biology | ||||||||||||||||
| Restrictions | Max. 40 studerents (lab facilities) | ||||||||||||||||
| Areas of Competence the Course Will Address | |||||||||||||||||
| Competences obtained within basic science: Comprehend the biology of a selection of relevant organisms representing common expression hosts. Transfer theory and principles regarding the usefulness of different organisms as expression hosts. Competences obtained within applied science: Comprehend the methods used to optimise the expression level and stabilise the product. Are able to apply principles to the expression of a given gene. Competences obtained within ethics and values: Reflects about the use of GM organisms for production of peptides. Reflects about the disease risks in connection to the choice of host. | |||||||||||||||||
| Course Objectives | |||||||||||||||||
| The production of technical enzymes as well as of peptide- and protein- based pharmaceuticals are in large scale being performed in specially designed host organisms. The aim of the course is to educate the students in processes associated with heterologous expression. The students will upon completion of this course be able to design and perform a strategy for the expression of a given gene. This includes considerations about amount, quality and downstream applications of the product. | |||||||||||||||||
| Course Contents | |||||||||||||||||
| The course will contain a theoretical part where most aspects of peptide and protein production in biological organisms will be discussed (see below). You will acquire knowledge on the variety of possible host organisms found in the different kingdoms. We will discuss the possibilities of designing and finding new suitable expression hosts. Topics that will be covered in the theoretical part of the course: The intelligent choice of a host organism / Cloning strategies envisioned by an "in silico" multistep cloning / Promoter strength and induction / Copy number and silencing problems in heterologous hosts / Expression vectors / mRNA stability and introns / Choice of, and placement of purification tags / Stability of the product / Secretion of proteins and signal trapping / Sub-cellular targeting and storage of the product / Post-translational modifications in different host organisms / Inclusion bodies and folding of proteins / Expression of membrane proteins compared to soluble proteins / Heterologous expression for production of antibodies / Expression of toxic proteins / Transient expression / Optimisation of expression level / Fermentation and large scale production. In the practical part of the course you will work with a range of different expression organisms. These include Escherichia coli, Saccharomyces serevisiae, Pichia pastoris, COS-cells, Xenopus oocytes and higher plants. You will transform all of them and determine the amount of produced protein, you will optimize the expression level and prove that the expressed protein exhibit the desired function. In the practical part we will not only focus on topics relevant for the particular organisms but also cover a broad aspect of typical problems related to the production of recombinant protein. An example is the requirement for a system where you can perform physiologically relevant measurements in order to test new drugs directed against ion-channels or receptors. Topics from the practical part of the course: Expression and assembly of a multi subunit protein complex / The effect of alcohol and temperature on expression level / Expression of a secreted protein / Sub-cellular fractionation / Detection of post-translated modifications / The use of protein homologs from thermophilic bacteria / Characterisation of antibodies against membrane proteins / Use of fluorescent markers for detection of expression level / Electrophysiological measurement on ion-channels / Receptor-ligand binding / Co-expression / Virus induced expression | |||||||||||||||||
| Teaching And Learning Methods | |||||||||||||||||
| The course contains both a theoretical part and a practical part. In the theoretical part there will be lectures as well as student presentations based on cases and journals. A laboratory course is running several days during most weeks. The course will be divided into smaller parts build upon the different expression organisms. | |||||||||||||||||
| Course Litterature | |||||||||||||||||
| The students will receive a collection of scientific papers together with the manuals for the laboratory part of the course. | |||||||||||||||||
| Course Coordinator | |||||||||||||||||
| Anja Thoe Fuglsang, atf@life.ku.dk, Department of Plant Biology/Plant Physiology and Anatomy Laboratory, Phone: 35333329 | |||||||||||||||||
| Study Board | |||||||||||||||||
| Study Committee NSN | |||||||||||||||||
| Course Scope | |||||||||||||||||
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