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Question 1

Two key features of a plasmid vector that you could use to generate corn plants that express GFP only in corn kettles

Answer 1

• promoter ( drive expression in corn Kernals)
• transgene (clone into plasmid )
• ori
• antibiotic resistance

Question 2

Why can’t proteins with complex post-translational modifications be produced in bacteria? What is al alternative protein production system that will allow successful expression of glycosyalted proteins?

Answer 2

• they dont have a golgi or ER
• they doing have enzymes required to glycosylation

Mammalian cells
Transgenic animals
Transgenic plants

Question 3

Worst and best choices of protein production

Answer 3

Question 4

In protein production ____ is a problem for bacteria

Answer 4

Folding

Question 5

Is insulin PTM and where is it best produced in?

Answer 5

No
Bacteria

Question 6

Is EPO PTM and where is it best produced in?

Answer 6

• yes, gycoslated
• mammalian cells (CHO)

Question 7

Is antithrombin PTM and where is it best produced in?

Answer 7

Gamma - carboxylated
(Not modified in mammalian cell) there for its best produced in transgenic animals (goats)

Question 8

GFP is usually found in

Answer 8

Jelly fish

Question 9

Insulin found in the

Answer 9

Pancreas

Question 10

Antithrombin is found in the

Answer 10

Kidney

Question 11

Purpose of protein evolution

Answer 11

• speeding up evolution process to try and alter or try and improve a protein for a new or better use

Question 12

What were the different colours of GFP made from

Answer 12

Protein evolution

Question 13

How protein evolution of GFP worked to make diff colours

Answer 13

• amplified the cDNA using degenerate PCR
• degenerate PCR allows for mutations as degerate doesnt have proof reading mechanisms, making it prone to errors
• take mutated PCR products and clone into plasmid (using restriction sites) then transform into bacteria.
• select out the variants from the plate
• can then sequence to discover what to base pair change was

Question 14

Recombinant DNA technologies in the clinic =

Answer 14

Recombinant proteins that require repeated administration of drugs for the life of the patient / illness

?????

Question 15

How can we provide a permanent treatment ?

Answer 15

Use patients as host for protein production

Question 16

Plasmids can be carries into proteins ALSO by using ____ _____

Answer 16

Viral vectors

Question 17

Viral and bacterial plasmids look the same

Answer 17

Yes

Question 18

In gene therapy we introduce a plasmid into a… which is then…….
Why?

Answer 18

…Viral vector (virus is diablesd) which is then inserted into a patient and gets taken up into cells and gene of interest is expressed under control of an appropriate promoter

• good at getting genetic info into cells

Question 19

Invivo vs exvico

Answer 19

Exvivo - take cells out of body to insert gene
Invivo - insert directly into cells

Question 20

Conventional biotechnology vs gene therapy

Answer 20

Question 21

What kind of promoter do we use for gene therapy insulin production in humans

Answer 21

Mammalian glucose responsive promoter

Question 22

Virus is used as a ____ to ___ ______ ____ patient _____

Answer 22

Virus is used as a vector to carry gene into patient cell

Question 23

In insulin gene therapy what cells of what organ are target and why

Answer 23

• liver rather then pancreas where cells are dying

Question 24

Which system is virus injected into in the permanent gene therapy hypothesis and what do they target

Answer 24

Venous
Liver - so they stay whole life time - only one treatment

Question 25

Why can’t u target pancrease B cells for gene therapy with insulin?

Answer 25

B cells are destroyed by autoantibodies so we use liver to make insulin (using promote that responds to glucose in the blood)

Question 26

Insulin - what kind of cDNA

Answer 26

- prepreprtein Fulll one

Question 27

Different protein factories are required depending on

Answer 27

Complexity of the protein u want to produce

Question 28

DNA technology can be used to..

Answer 28

Create new proteins

Question 29

Using humans as a protein factory (gene therapy) can be used to…

Answer 29

Permanently treat disease