Recombinant proteins Flashcards

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1
Q

Give 3 examples of therapeutic recombinant proteins.

A
  1. Hormones like insulin
  2. Vaccines
  3. Antibodies
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2
Q

What are diagnostic proteins?

A

Those used to determine an outcome.

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3
Q

Give 3 examples of diagnostic proteins.

A
  1. Enzymes
  2. Antibodies
  3. Biosensors
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4
Q

Commercial enzymes can be used in a) industry and b) biotechnology. Give examples of each.

A

a) Food production

b) Restriction enzymes, thermostable polymerases etc.

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5
Q

In order to recombine proteins you must first obtain pure forms of those you wish to combine. Can you get these from natural sources?

A

Yes if they are abundant in a particular cell or tissue.

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6
Q

What are 3 disadvantages to purifying proteins from natural sources?

A
  1. Cost
  2. Ethical considerations
  3. Contamination/infection
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7
Q

You can obtain proteins for recombination from a transformed bacterial host instead. Give 5 advantages of this.

A
  1. Produces large volumes of protein by clonal growth
  2. Genetic engineering allows for high expression of required protein
  3. Reduced cost
  4. Reduced risk of contamination
  5. More ethically acceptable
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8
Q

If you are transforming bacteria with eukaryotic DNA what must you use and why?

A

cDNA because it contains no introns.

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9
Q

Promoters are usually organism-specific and thus un-transferable. True or false?

A

True, for example a human promoter will not work in yeast.

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10
Q

You must always use endogenous promoters in protein production. These are not only organism specific but tissue specific as well. True or false?

A

True.

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11
Q

The genetic code is degenerate. What does this mean?

A

Multiple codons code for the same amino acid.

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12
Q

Different organisms display preference for some codons. True or false?

A

True - because the code is degenerate several codons produce the same AA, and some species have higher proportions of particular codons.

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13
Q

What is codon optimisation?

A

Adapting the genetic code to suit the host organism, e.g. using codons that they prefer.

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14
Q

Give 4 methods of delivery of synthetic genes into a host.

A
  1. Electroporation
  2. Chemical treatment
  3. Biolistics
  4. Microinjection
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15
Q

Define electroporation.

A

Delivering a pulse of electricity that momentarily opens the pores of a cell.

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16
Q

Define biolistics.

A

Whereby tiny metal pellets coated in DNA are propelled into living cells at high velocities.

17
Q

The expression of a protein in a host does not always work as it creates negative selection against the host. True or false?

A

True.

18
Q

Inserted proteins may not always be expressed in the host. Give 3 reasons why.

A
  1. Point mutations
  2. Methylation
  3. Chromosomal rearrangements
19
Q

If the inserted protein is negatively selected for, why might it not be expressed?

A

Because it is unfavourable.

20
Q

Expression can go wrong and result in a) degradation and b) aggregation. What does this mean?

A

a) Where the protein is broken down and becomes non-functional
b) Too much is produced and clumps together, also reducing function

21
Q

Where is hGH produced?

A

In the pituitary gland.

22
Q

Animal hGH is compatible with humans. True or false?

A

False - it was isolated from human cadavers but there were major issues with HIV and prion proteins and this is now banned.

23
Q

How is hGH produced now?

A

With E. coli as a host.