Expression

Question 1

Steps - Expression

Answer 1

• Add gene of interest into a host system
• Select gene of interest & grow large quantites of cells
  -Collect cells for purification

Question 2

Parts Plasmids

Answer 2

• Promoter (RNA Pol binds)
• Multiple Cloning site (gene of interest inserted)
• ORI Replication start site
• Antibiotic resistance gene
• Selectable mrker
• Solubility tag

Question 3

4 host systems

Answer 3

Bacteria
Yeast
Insect
Mammalian

Question 4

Bacteria host system

Answer 4

+
Less expensive
High protein yields
Rapid expression (1 week)
Easy to scale up

-
No PTM
Few peoteins are secreted
Some proteins undergo misfolding/aggregration

Question 5

Yeast Host System

Answer 5

+
Less expensive
Medium-high protein yields
Moderately rapid expression
Some PTM
Some secreted proteins

-
Some protein structures are different than mammalian counterpart
Requires yeast secretion peptides

Question 6

Insect Host System

Answer 6

+
Moderately rapid expression
Most PTM
Secreted and membrane proteins
High protein yields

-
Some proteims structures are different than mammalian counterpart
Expensive
Use of baculovirus

Question 7

Mammalian Host system

Answer 7

+
PTM
Good for secreted nd membrane proteins

-
Expensive
Loew protein yields
Expression takes months (2-3)