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Flashcards in Perfect and problematic expression Deck (5)
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1

Why is there a protein solubility problem in E. coli?

The dissimilarity of eukaryotic proteins and E. coli post-translational processes

2

What can't E. coli do to a protein?

Glycosylation

3

What happens to the proteins produced?

They are produced as insoluble deposits in the cell

4

Describe denaturation:renaturation

Add denaturants to form linear polypeptides
Remove denaturants to refold into soluble proteins

5

How to avoid the protein folding problem?

Express the protein at a lower temperature
Express the protein at a lower level to avoid the aggregate formation
Target the protein to the periplasm as folding and disulphide interchange occurs more readily in this compartment
Express protein as a fusion with GST or thioredoxin to enhance disulphide bond interchange and folding in the cytoplasm
Use host strain for expression which overexpresses chaperonins
Use eukaryotic host instead