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BIOC192 > Q/A Session 4/5 > Flashcards

Q/A Session 4/5 Flashcards

1
Q

What is a receptor?

A

Protein molecule that receives chemical signals from outside the cell

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2
Q

What is a ligand?

A

Molecule/drug that binds to a receptor

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3
Q

When alcohol binds to the GABAa receptor what enters the cell?

A

Cl- ions into the cell

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4
Q

What is the role of HIV protease?

A

Cleave long proteins into fragements

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5
Q

What kind of inhibitor will a pharma drug be?

A

A competitive inhibitor

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6
Q

What do transcriptional factors do?

A

Determines when and in what cell a gene is transcribed to produce mRNA

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7
Q

What are transcription factors?

A

Proteins that bind to a specific DNA sequence and control rate of transcription

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8
Q

What is a promoter?

A

DNA sequence at which transcription factors bind and recruit RNA polymerase

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9
Q

What does a promoter determine?

A

If and how much a gene is transcribed

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10
Q

What 4 things does PCR require?

A
  1. DNA from cell
  2. DNA from nucleotides
  3. Primers with sequence that can base pair to region of amplification
  4. Taq DNA polymerase
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11
Q

What are the 3 steps of PCR

A
  1. Heat DNA to 95 degrees to separate DNA strands. Heat breaks the H-bond
  2. Cool to ~60 degrees to base pair a DNA primer. Primer=short DNA sequence of nucleotides complementary to strand
  3. Heat DNA to 72 degrees to allow Taq DNA polymerase to copy DNA.
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12
Q

What do tumour suppressor genes do?

A

Encode proteins that prevent uncontrolled cell growth

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13
Q

What do onco-genes do?

A

Promote cell division

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14
Q

What is a proto-oncogene?

A

A normal gene that when mutated promotes uncontrolled cell growth

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15
Q

What are the components of recomb DNA plasmids?

A
  1. Origin of Replication
    - Allows initiation of replication using host DNA polymerase
  2. Antibiotic Resistance gene
    - Allows selection of cells containing plasmid
  3. Promoter
    - Drives expression of gene in cells with appropriate transcription factor machinery
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16
Q

What are the characteristics of plasmids?

A
  • Circular pieces of dsDNA
  • Replicate independently of host chromosomal DNA, but use host machinery
  • Provide a benefit to host
17
Q

What are restriction enzymes?

A
  1. Cut dsDNA into specific sequences creating sticky ends

2. Found naturally in bacteria-degrade foreign DNA

18
Q

What is transformation?

A

Transfer of plasmids into bacteria

19
Q

What are the 5 steps of producing recombinant protein in prokaryotes?

A
  1. Isolate gene of interest
  2. Clone into expression plasmid
  3. Transform bacteria for expression
  4. Grow cells expressing gene of interest
  5. Isolate and purify protein
20
Q

Advantages of prokaryote systems

A
  • Low cost
  • high yield
  • pathogen free
21
Q

Disadvantages of prokaryote systems

A
  • Proteins often partially folded

- Inability to perform post-translational modifications

22
Q

Advantages of mammalian cell recombs

A
  • Protein can be produced as pre-pro-protein and processed efficiently
  • Secreted from cells: easier purification
23
Q

Disadvantages of mammillian systems

A

More expensive

24
Q

What are the 5 steps of producing recombinant protein in eukaryotes?

A
  1. Isolate cDNA
  2. Clone into eukaryotic expression plasmid
  3. Transform bacteria to produce more plasmid DNA and then transfect eukaryotic cells
  4. Extract recomb from cell media.
  5. Purify
25
Q

What is glycosylation?

A

Requires mammalian cells. Adding carbohydrates to residues

26
Q

What is pharming?

A

Using whole animals to make recomb proteins

27
Q

What is insulin best produced in?

A

Bacteria. A+B chains are produced separately.

The PTM is formation of disulfide bonds

28
Q

What is EPO best produced in?

A

Mammalian cell culture. Hamster Ovaries

PTM is glycosylation

29
Q

What is Antithrombin best produced in?

A

Transgenic animals

PTM is carboxylation

30
Q

What does gene therapy use?

A

Viral vector for stable integration into host cell

->gene encoding recomb protein

BIOC192 (4 decks)

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