Perfect and problematic expression Flashcards Preview

BIO2030 Biotechnology > Perfect and problematic expression > Flashcards

Flashcards in Perfect and problematic expression Deck (5)
Loading flashcards...
1
Q

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

A

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

2
Q

What can’t E. coli do to a protein?

A

Glycosylation

3
Q

What happens to the proteins produced?

A

They are produced as insoluble deposits in the cell

4
Q

Describe denaturation:renaturation

A

Add denaturants to form linear polypeptides

Remove denaturants to refold into soluble proteins

5
Q

How to avoid the protein folding problem?

A

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