Trygve Brautaset - Recombinant Expression/Protein Production Flashcards
A heterologous gene originating from a plant is to be expressed recombinant in the bacterium Escherichia coli.
a. Describe genetic modifications needed to achieve this.
b. You manage a) but you observe very poor expression of your heterologous gene product; describe three different strategies you can test to increase expression level.
c. Your gene product is host-toxic; describe two different strategies that can be done to overcome this problem.
Fusion technology is a technique with many different applications; describe how this technique can be used to solve problems related to:
a. Protein solubility
b. Protein purification
c. Protein detection
Plasmids are commonly used as expression vectors; describe similarities and differences between the pET and pBAD plasmid vectors for induced recombinant expression in Escherichia coli.
Describe how mRNA stability of one particular gene can be affected by translation efficiency of this gene in Escherichia coli.
Industry use fermentors to scale up recombinant protein production. Describe advantages and challenges by cultivating recombinant E. coli in fermentors and describe why it is particularly useful to use inducible expression systems in such reactors.
