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Biology A2 Unit 2 > Gene Technology > Flashcards

Gene Technology Flashcards

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1
Q

What are GMO’s

A

Organisms with recombinant DNA

Can be split into genetically engineered microorganisms (GEM’s) and others

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2
Q

What are the containment methods for GEM’s

A

⭕️ bacterial strains ill-adapted to the human physiology - minimum temp tolerance above human body temp
⭕️ contain ‘suicide genes’ - activated if move outside certain pH or temp range
⭕️ purpose-built licensed laboratories
⭕️ tightly controlled procedures for staff - cleaning etc

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3
Q

What are the three methods of genetically modifying a plant

A
  1. Use a tumour inducing plasmid from the common soil bacterium
  2. Use gene guns
  3. Use plant viruses
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4
Q

Describe how you would use a tumour inducing plasmid to genetically modify a plant

A
  • Agrobacterium tumefaciens invades damaged plant tissue
  • tumour-inducing (Ti) plasmid transferred to plant cell
  • desired gene spliced into plasmid
  • readily taken up by plant cell provided cell wall has been removed by enzyme cellulase
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5
Q

Explain how gene guns are used to genetically modify a plant

A
  • minute pellets covered in DNA w desired gene
  • shot through cellulose walls
  • using gene gun
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6
Q

What are the advantages of GM crops

A

See spider diagram in notes

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7
Q

What are the disadvantages of GM crops

A

See spider diagram notes

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8
Q

How are plant viruses used to genetically modify a plant

A

Infect cells by inserting their nuclei acid

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9
Q

Describe how to extract and cut up DNA

A
  • restriction enzymes - cut by hydrolysis
  • recognition sequence
  • leaves sticky ends
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10
Q

Describe a sticky end

A

Short single stranded section of DNA

Bases are unpaired so exposed

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11
Q

What does reverse transcriptase do

A

Uses mRNA as template strand
Produces single stranded cDNA
Which is converted to double stranded DNA by DNA polymerase

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12
Q

What is a gene probe and what do they do

A

Short single strand of DNA
Binds to target gene
Confirm presence of desired gene
Identify sections of DNA

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13
Q

What are the steps for using a fluorescently labelled probe

A
  1. DNA cut
  2. Separate by gel electrophoresis
  3. Transferred to nylon membrane
  4. Treated w fluorescently labelled probe
  5. Probe binds
  6. Exposed to UV light
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14
Q

What is pcr

A

Few fragments of DNA duplicated to millions

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15
Q

What enzyme is used in pcr

A

Taq polymerase

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16
Q

What is the function of primers

A
  • stop two DNA strands rejoining
  • ‘bracket’ section of DNA to be copied
  • DNA replication can only start with a double stranded region
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17
Q

What is the process of PCR

A
  1. Heat to 95°C - break H and separate
  2. Cooled to 50-60°C - primers annealed
  3. Heat to 72 °C -enzyme optimum temp
  4. Repeat
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18
Q

What is the role of PCR

A

Forensic analysis
Pre natal diagnosis
Historical studies
Genetic relationships

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19
Q

What are MRSs

A
  • microsatellite repeat sequence
  • Small no. of bases repeated many times in non-coding region
  • number of repeats is unique to each individual
  • probe for particular MRSs created
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20
Q

What are SNPs

A

Differences in single nucleotides

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21
Q

What are the steps for genetic fingerprinting

A
  1. Extract DNA
  2. Restriction enzymes cut out repeat sequence
  3. Separate fragments by gel electrophoresis
  4. Transfer to nylon sheet - southern blotting
  5. Attached labelled probe
  6. detect diff repeat sequence
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22
Q

What is the role of genetic fingerprinting

A 
Establish paternity
Forensics
Tissue typing for transplants
Evolutionary relationships 
Identify victims
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23
Q

What is a vector

A

Length of DNA

carries gene we want into host cell

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24
Q

What is a plasmid

A

Small circular ring of DNA
Bacterial cell
Copy themselves independently

25
Q

What are the two methods of obtaining required gene

A
  1. Restriction endonuclease cut gene out of chromosomal DNA

2. Reverse transcriptase - produce DNA from RNA

26
Q

What are the two methods for inserting gene into vector

A

Bacterial plasmids

Viruses

27
Q

How are bacterial plasmids used to insert gene into a vector

A
  1. Plasmid cut w restriction enzyme
  2. Same enzyme cuts gene from donor DNA
  3. Gene spliced into plasmid
  4. Sticky ends anneal
  5. DNA ligase - phosphodiester bonds between sugar-phosphate backbones
  6. Now recombinant plasmid
28
Q

How are viruses used to insert gene into a vector

A
  1. Insert genetic material into host cell

2. Transfer recombinant DNA into bacterial cell

29
Q

What are the two methods for inserting the vector into a host cell

A

Heat shock

Electroporation

30
Q

Describe the method of heat shock

A
  1. Cells incubated w recombinant plasmid
  2. In solution of calcium ions at 0°C
  3. Sudden rise in temp to 40°C
  4. Bacterial cells more permeable fo plasmids
31
Q

Describe electroporation

A
  1. Short electrical pulse
  2. Temporarily disrupts membrane
  3. Opens pores in membrane
32
Q

Describe how a marker gene is used to check cell contains recombinant DNA

A

⭕️ R-plasmid resistant to tetracycline and ampicillin
⭕️ restriction enzyme cuts tetracycline gene but not ampicillin
⭕️ if it has taken up original plasmid then it’s resistant to both
⭕️ if takes up recombinant plasmid then resistant to ampicillin but not tetracycline

33
Q

How to use fluorescent protein to check if cell contains recombinant DNA

A

Gene that produces fluorescent proteins isolated and attached to gene of interest
If taken up then will glow under UV light

34
Q

How to make a replica plate

A
  1. Sample on master plate
  2. Touch cloth on bacterial colonies on master plate
  3. Touch on to replica plate containing tetracycline
35
Q

How to clone transformed bacteria

A

Cultured in fermenters
Transferred gene switched on
Optimal pH, temp and nutrients

36
Q

What are the 4 methods of introducing genes into animal cells

A

Electroporation
Liposomes
Viruses
Microinjection

37
Q

Describe how liposomes work when inserting genes into animal cells

A

Coat DNA in artificial lipid vesicles

Adhere to cell membrane - pass DNA into cell

38
Q

What are the two types of viruses used to introduce genes into cells

A

Adenoviruses

Retroviruses

39
Q

How do retroviruses work

A

RNA delivered to host
Copied to DNA
By reverse transcriptase

40
Q

How do microinjections work when introducing genes to animal cells

A

Inject DNA directly into nucleus of fertilised egg

41
Q

List some of the uses of genetically modified animals

A

Protein enriched milk - cow
Treatment of diseases
Treatment of skin cancer - chicken eggs

42
Q

What are the two types of gene therapy

A

Somatic cell gene therapy

Germ line therapy

43
Q

What are the pros and cons of somatic gene therapy

A

C - only targets affected tissue
C - only when defective gene in specific and easily reached part of the body
P - used at any stage of individuals life

44
Q

What happens when functional allele is in host cell

A
  1. Donor DNA incorporated in host DNA

2. DNA functions as independent DNA unit (episome)

45
Q

What is a genome

A

Complete nucleotide (base) sequence

46
Q

What is genome sequencing

A

Determining order of bases in each chromosome of eukaryotic organisms

47
Q

What is genetic mapping

A

Establishing exactly where particular genes are on chromosomes

48
Q

What is a microarray

A 
⭕️ solid silicon/glass base
⭕️ thousands of spots 
⭕️ spots contain many of same DNA probe
⭕️ diff probes in diff spots
⭕️ analyse many genes simultaneously 
⭕️ rapid analysis of DNA
49
Q

Why use microarays

A
  1. Identify genotypes - detect defective alleles - diagnose genetic disease
  2. Measure gene expression - help diagnose cancer - gene more or less active
50
Q

How is a microarray used

A

⭕️ single stranded DNA/mRNA washed over array
⭕️ complementary to probe the hybridise with it
⭕️ DNA/mRNA labelled
⭕️ diff colours = diff levels of hybridisation

51
Q

How is gene expression measured using microarrays

A
  1. mRNA extracted - concerted or cDNA by reverse transcription
  2. Flourescent tag
  3. Added to array
  4. Complementary then will hybridise
52
Q

How can a microarray be used to diagnose oncogenes

A
  1. Labelled cDNA from RNA from healthy and cancerous tissue
  2. Dif flourescent dyes for each
  3. Both added to microarray and hybridise
  4. Scan with laser and tags generate signals - strength of which depends on amount of cDNA binding to probes
53
Q

What are the benefits of genome sequencing

A
  • mapping of genes
  • pharmacogenetics - designer drugs
  • gene therapy
  • DNA fingerprinting
  • ancestral links
  • work out primary structure of protein
54
Q

What is a designer drug

A
  • Matched to individuals profile
  • more effective first time
  • reduce trial and error so less suffering
  • reduce side effects and allergies
55
Q

What is gene knockout

A

Gene(s) removed or made inoperative

Allow you to determine the role of the gene

56
Q

What is gene knockin

A

Particular gene added

Deliberately add defective gene to study the disease progression

57
Q

Why are mice used in studies

A

Closely related to humans
Mammal
Short life cycle and easily kept in lab conditions
Most people not ethically opposed

58
Q

Pros and cons of genetic screening

A

✅ understand risk of passing on genetic disorder
❌ stress if carrier
❌ termination of pregnancy?

Biology A2 Unit 2 (9 decks)

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