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BIOC192 > 21 > Flashcards

21 Flashcards

1
Q

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

A
  • promoter ( drive expression in corn Kernals)
  • transgene (clone into plasmid )
  • ori
  • antibiotic resistance
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2
Q

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?

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

Mammalian cells
Transgenic animals
Transgenic plants

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3
Q

Worst and best choices of protein production

A
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4
Q

In protein production ____ is a problem for bacteria

A

Folding

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5
Q

Is insulin PTM and where is it best produced in?

A

No
Bacteria

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6
Q

Is EPO PTM and where is it best produced in?

A
  • yes, gycoslated
  • mammalian cells (CHO)
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7
Q

Is antithrombin PTM and where is it best produced in?

A

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

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8
Q

GFP is usually found in

A

Jelly fish

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9
Q

Insulin found in the

A

Pancreas

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10
Q

Antithrombin is found in the

A

Kidney

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11
Q

Purpose of protein evolution

A
  • speeding up evolution process to try and alter or try and improve a protein for a new or better use
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12
Q

What were the different colours of GFP made from

A

Protein evolution

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13
Q

How protein evolution of GFP worked to make diff colours

A
  • 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
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14
Q

Recombinant DNA technologies in the clinic =

A

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

?????

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15
Q

How can we provide a permanent treatment ?

A

Use patients as host for protein production

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16
Q

Plasmids can be carries into proteins ALSO by using ____ _____

A

Viral vectors

17
Q

Viral and bacterial plasmids look the same

A

Yes

18
Q

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

A

…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
19
Q

Invivo vs exvico

A

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

20
Q

Conventional biotechnology vs gene therapy

A
21
Q

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

A

Mammalian glucose responsive promoter

22
Q

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

A

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

23
Q

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

A
  • liver rather then pancreas where cells are dying
24
Q

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

A

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

25
Q

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

A

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

26
Q

Insulin - what kind of cDNA

A
  • prepreprtein
    Fulll one
27
Q

Different protein factories are required depending on

A

Complexity of the protein u want to produce

28
Q

DNA technology can be used to..

A

Create new proteins

29
Q

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

A

Permanently treat disease

BIOC192 (38 decks)

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