240045 Heterologous Expression, Theoretical

Details
Department of Plant Biology   85 %
Department of Basic Animal and Veterinary Sciences   10 %
Department of Veterinary Pathobiology   5 %
Earliest Possible YearMSc. 1 year to MSc. 2 year
DurationOne block
 
Credits7.5 (ECTS)
Course LevelMSc
 
ExaminationFinal Examination

oral examination


All aids allowed

Description of Examination: The exam will be based on a number of selected cases (15) among the ones discussed during the course. The text book is considered as background.

Weight: The final exam count 100%



13-point scale, internal examiner
 
Requirement For Attending Exam75% of the written cases must be approved in order to attend the exam.
 
Organisation of Teaching
 
Block PlacementBlock 3
Week Structure: A
 
Teaching LanguageEnglish
 
Optional Prerequisites240042 Thematic Course: Experimental Molecular Biology
230003 Biochemistry 2
 
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 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 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.
 
Teaching And Learning Methods
This course is a theoretical course. There will be lectures as well as student presentations based on cases and journals. The course will be divided into smaller parts build upon the different expression organisms. The students must write a small case (2 pages) every week.
 
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
lectures24
Colloquia16
Excursions10
preparation95
project work60
examination1

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