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George Biology > Manipulating genomes chp 21 > Flashcards

Manipulating genomes chp 21 Flashcards

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1
Q

define genetic engineering

A

manipulation of the genome

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2
Q

define genome

A

all the genetic information found within an organism

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3
Q

define transgenic

A

organism that carriers a gene from another organism

from another species

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4
Q

what type of enzymes are used in genetic engineering

A
  • restriction endonuclease
  • DNA ligase
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5
Q

what is the function of restriction endonuclease

A
  • cuts required gene from DNA
  • cuts open DNA in host organism
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6
Q

define sticky ends and why are they made

A
  • regions with unpaired, exposed bases
  • many restrictions endonuclease cut the DNA strand unevenly, leaving strand fragmented
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7
Q

why do we want sticky ends to be produced during genetic engineering

A

make it easier to insert desired gene into the DNA

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8
Q

how can mRNA be used for genetic engineering

A
  • isolate mRNA for desired gene and using enzyme reverse transcriptase to produce strand of complementary DNA
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9
Q

what enzyme is used in genetic engineering involving mRNA

A

reverse transcriptase

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10
Q

what advantages are there of using genetic engineering involving mRNA

A
  • its easier to identify desired gene, as particular cells will make some very specific types of mRNA
    ^beta cells in pancreas make insulin mRNA
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11
Q

define recognition site

A

genetic sequences where the restriction enzymes cut the DNA segments

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12
Q

define vector in genetic engineering

A

something that carries the isolatyed DNA into the host cell

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13
Q

what are some common used vectors in genetic engineering

A
  • plasmid DNA
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14
Q

define plasmid DNA

A

small circular molecules of DNA separate from chromosomal DNA that can replicate independently

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15
Q

define recombinant DNA

A

when isolated DNA combines with host DNA

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16
Q

what Is the function of DNA ligase in genetic engineering

A

forms phosphodiester bonds between the sugar and phosphate groups on two strands of DNA

17
Q

define marker gene

A

a gene used to determine if DNA sequence has been successfully inserted into an organism’s DNA

18
Q

How can marker genes be used to test if the gene insertion has been successful

A
  • have characteristic that can be used to see if insertion done correct
  • common one is antibiotic resistance, bacteria then grown in antibiotics (if survive then done correctly)
19
Q

define transformation in genetic engineering

A

process of transferring recombinant DNA into host cell

20
Q

define electroporation

A
  • small electrical current applied to bacteria
    ^ this makes membrane porous so plasmids can move into cell
21
Q

what is a problem with electroporation

A

if too much electrical current used can destroy whole cell

22
Q

define electrofusion

A
  • tiny electric currents are applied to membranes of 2 different cells
    ^this fuses cell and nuclear membranes to form polypoid cell
23
Q

what is electrofusion used for

A
  • producing monoclonal antibodies
24
Q

what are genetically engineered prokaryotes used for

A

produce:
- hormones (insulin, HGH)
- clotting factors
- antibiotics
- pure vaccines

25
Q

what are the pros of genetic engineering in plants

A
  • drought resistance
  • higher yeilds
  • producing herbicides & pesticides (so dont need to be sprayed)
  • can improve nuitritional value
  • reduces economic costs and carbon foot print
26
Q

what are some cons of genetically engineered plants

A
  • could have adverse side effects
  • could have unknown effects
  • may limit biodiversity as out compete local wildlife
  • cross pollunation could lead to superweeds
  • patents can create monopolies (high prices)
27
Q

what form of transformation is used to genetically engineer plants

A
  • electrofusion
28
Q

why is it hardest to genetically engineer eukaryotic animals cells in particular

A
  • cell membranes are hard to manipulate
29
Q

what are the steps to genetically modify plant cells

A
  • cut lear
  • expose cells to bacteria carrying desired gene and marker gene (antibiotic resistance)
    ^allow bacteria to deliver genes
  • expose leafs to antibiotics
  • ones that survive have desired gene, let them multiple & form clump
  • allow callous to grow shoots + roots
  • transfer plant to soil
30
Q
  • what percentage of DNA codes for proteins
A
  • 2%
31
Q
  • what is non-coding DNA called
  • What is coding DNA called
A
  • introns
  • exons
32
Q

what are some uses for DNA profilling

A
  • paternity test
  • forensic identification at crime scenes
  • Diagnosing genetic diseases
33
Q

what are the 2 sub-groups within non-coding DNA

A
  • VNTR (variable number tandem repeats ) also called minisatillites
  • STR (short tandem repeats) also called microsatilistes
34
Q
  • What type of DNA is used for DNA profilling
  • why is this
A
  • introns as exons
  • using coding sequence of DNA would not provide unique proflies
35
Q

what are the stages of DNA profilling

A
  • extract DNA
  • amplify it using PCR (artifical DNA replication)
  • cut DNA using restriction endonuclease
  • place DNA into wells on gel electrophoresis plate
    ^seperate DNA based on mass
  • Dye DNA using fluroescent/radioactive dye and view under X-ray/UV
36
Q

what does PCR do

A
  • makes more DNA
37
Q

Explain process of how PCR works

A
  • heat up DNA to break hydrogen bonds
  • cool solution slightly so can add primer
  • DNA polymerase (from bacteria in hot climates) used to speed up PCR
    ^solution heated to optimum temp for this
38
Q

What is a primer in PCR

A
  • short sequence of DNA bases that are complmentary and specific to part of DNA you want to copy

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