genetic engineering Flashcards

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1
Q

state the 5 steps of DNA technology (in vivo cloning)

A
  • create DNA fragment
  • insert DNA into a vector
  • transform host cell with vector
  • identify transformed cell
  • grow host cell (clone)
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2
Q

how is DNA amplified?

A
  • using PCR
  • cDNA into thermal cycler
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3
Q

summarise how bacteria is used to produce human insulin

A
  • mRNA extracted from cells and treated with reverse transcriptase to make complementary DNA/plasmid obtained from bacteria & cut with restriction enzyme
  • plasmid and cDNA fuse using DNA ligase
  • recombinant plasmid introduced into host cells
  • bacteria multiply in a fermenter and produce insulin
  • separation & purification of human insulin can be used by diabetic patients
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4
Q

what are the advantages of using bacteria in the process?

A
  • multiply quickly
  • DNA is easily accessible (has free/plasmid DNA in cytoplasm)
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5
Q

describe how a plasmid is cut

A
  • both strands of DNA cut
  • using a specific restriction enzyme
  • in a particular section
  • creates sticky ends
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6
Q

how can the same sticky end be created?

A

use same restriction enzyme - will cut at same recognition site

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7
Q

why are sticky ends made to be identical?

A

cut plasmid & cDNA align and the cDNA becomes part of the plasmid

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8
Q

how can the plasmid be inserted into a bacterium?

A
  • injected (using a vector)
  • use a solvent / detergent
  • increase temperature
  • electric shock bacteria cell
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9
Q

how can bacteria that have taken up insulin be detected?

A

inserting an indicator / antibiotic resistant bacteria when cDNA is inserted to show those that have picked up plasmids

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10
Q

what is a plasmid?

A

a small, circular, double-stranded DNA molecule that naturally exist in bacterial cells - often provide bacteria with antibiotic resistance.

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11
Q

what is cDNA

A

copy DNA

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