Gene Tech Flashcards

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1
Q

Define genome

A

Entire set of dna

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2
Q

What form does the dna need to be in to sequence the genome

A

Split into dna fragments

Then put in the right order

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3
Q

Define proteome

A

All the proteins an org can make

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4
Q

Why is the proteome of prokaryotes easy to find

A

Don’t have much non coding dna

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5
Q

Why is prokaryote proteome sequencing useful

A

Identifying antigens on virusues or bacteria to make medication and vaccines

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6
Q

Why is it harder to sequence the proteome of eukaryotes

A

Lots of non coding dna

And regulatory genes

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7
Q

What are regulatory genes

A

Genes that tell other genes to be switched on or off

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8
Q

What is recombinant dna technology

A

Transferring a fragment of dna from org to another

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9
Q

Why can dna from org be transferred to another org in a different species

A

Dna is universal
So it codes for the same amino acids

Protein synthesis mechanism are similar as well

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10
Q

What are orgs that have had transferred dna inserted into them

A

Transgenic organisms

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11
Q

Why is mRNA of a gene easier to get than the actual dna

A

Most cells only have 2 copies of the gene in dna

But lots of copies of the gene in mRNA form

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12
Q

What is reverse transcriptase used for

A

mRNA -> cDNA (complementary dna)

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13
Q

What is the method for productible dna fragments using reverse transcriptase

A

mRNA isolated from cells
Mixed with free dna nucleotides
Mixed with reverse transcriptase
mRNA acts as a template to make cDNA

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14
Q

What does restriction endonuclease recognise

A

Palindromic sequences/recognition sequences

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15
Q

What are palindromic sequences

A

Antiparallel pairs that read the same in opp directions

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16
Q

What is function of restriction endonuclease

A

Cut dna

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17
Q

Why does specific restriction endonuclease cut specific sections of dna

A

The shape of the recognition sequence is complimentary to the as

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18
Q

How do u get a section of dna between 2 palindromic sequences

A

Incubate restriction endonuclease to cut the palindromic sequences
Leaving the dna fragment

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19
Q

What type of reaction happens when dna is cut by restriction endonuclease

A

Hydrolysis

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20
Q

What is left by restriction endonuclease

A

Sticky ends

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21
Q

What are sticky ends

A

Tails on unpaired dna bases at the end of the fragment

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22
Q

What can sticky ends be used for

A

Anneal to a fragment with the complementary sticky end

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23
Q

What is the gene machine

A

Make dna from scratch without a preexisting template

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24
Q

How does the gene machine work

A

Sequence designed
First nucleotide of the sequence is attracted to a support (bead)
Nuceloutudes added in the correct order with protecting groups
Forming oligonucleotides
Broken off from support
Oligonucleotides are joined together to make a longer dna fragment

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25
Q

What is the purpose of protecting groups

A

Stop unwanted branching

Make sure the nucleotides join at the right points

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26
Q

What does in Vivo mean

A

inside a living org

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27
Q

What does amplifying dna mean

A

Make lots of copies

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28
Q

Describe the method of amplifying dna in Vivo

A
  1. Dna fragment inserted into a vector
  2. Vector dna cut open using the same restriction endonuclease that was used to isolate the dna
  3. Forms complementary sticky ends to the dna fragment
  4. Vector dna and fragments are mixed with dna ligase
  5. Vector dna and dna fragment join together = recombinant dna
  6. Host cells take up the gene (become transformed)
  7. Marker genes are inserted into the vector at the same time the dna fragment is added
  8. Host cells grow on an agar plate. Any colony formed from a transformed cell will all have the recombinant dna and marker gene
  9. Identify transformed cells eg. Uv light
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29
Q

What is ligase used for

A

Joins sticky ends of dna fragment to sticky ends of vector dna

=ligation

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30
Q

How do host cell cells take up recombinant dna from a plasmid vector

A

Host cells placed in ice cold CaCl2 = cell walls more permeable
Plasmids added
Mixture is heat shocked to get the plasmids to be taken up

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31
Q

How is recombinant dna taken up by host cells if the vector is a bacteriophage

A

The bacteriophage will inject its dna into the host cell

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32
Q

How do u produce proteins from the dna fragment

A

Vector needs to have specific promotors and terminator regions

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33
Q

What are promoter regions

A

Tell rna polymerase when to start productions mRNA

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34
Q

What does in vitro mean

A

Outside of living organisms

35
Q

What is the process used in vitro dna amplifying

A

Pcr

Polymerase chain reaction

36
Q

Décor cuve the method of in vitro dna amplifying

A
  1. Reaction mixture = dna fragment, free nucleotides, primers, dn a polymerase
  2. Heated to 95 = break h bonds between bases
  3. Cooled to allow primers to anneal to the strands
  4. Reaction heated to 72
  5. DNA polymerase lines up free nucleotides for complementary bases
  6. 2 new strands are formed
37
Q

What is a primer

A

Short piece of dna that complementary to the bases at the start of the dna fragment

38
Q

How many strands are made from each cycle of pcr

A

Double

39
Q

What does DNA polymerase do

A

Lines up free nucleotides for complementary bases

40
Q

How do u get plants to produce a desirable protein

A
Make a fragment of dna for the desired gene
Insert into plasmid
Insert plasmid into bacteria 
Insert bacterial dna into plant cell
Add the right promoter gene
41
Q

How do u make w genetically modified animal

A

Get gene that codes for a desirable protein
Inserted into early embryo or female egg cell
All cells dividing from the embryo will contain the new gene

42
Q

How are promoter regions specific

A

Only activated in specific cell types

Can be used to control the which cells the gene is expressed in

43
Q

Why are promoter regions so specific

A

Protein harvested more easily

If it was produced in the wrong cell it would be damaging

44
Q

How is gm used in agriculture

A

Higher yields = stop famine
More nutrients = stop malnourishment
Pest resistance = fewer pesticides needed
Reduce the costs to farmers

45
Q

How is gm used in industry

A

Use biological catalysts
Produced in large quantities
Less money

46
Q

How is gym used in médecine

A

Make vaccines and drugs quickly and cheaply

Make more efficient médecines = human insulin instead of pigs

47
Q

How is gm in agriculture bad

A

Monocultures = whole crop vulnerable to the same disease because they’re all genetically identical
Reducing biodiversity
Supersedes = gm plants interbreed with weeds = resistant to herbicides
Uncontrolled spread of recombinant dna in the ecosystem
Contaminate organic farmers = loose income

48
Q

How is gm used in industry bad

A
  1. Anti globalisation = a few large companies control the genetics industry. Force small businesses to close
  2. Without proper labelling = people won’t know what they are eating
  3. Loss of money as some consumer markets won’t buy gm
49
Q

How is gym used in médecine bad

A

Companies who own tech may limit its use = but it could be saving lives

Could be used unethically = designer babies

50
Q

What are the ethical issues of gm

A

Ownnpership issues
Does the donator or the scientists who modifies the dna own it
Patent seeds = High prices for gym seeds that farmers have to buy every year legally

51
Q

Name 4 benefits to gm for humans

A
  1. Drought resistant and better nutrition in crops
  2. Vaccines more accessible eg. In place where it can’t be refrigerated
  3. Médecines produced more cheaply
  4. Gene therapy to treat genetic diseases
52
Q

What is gene therapy used for

A

Treating human diseases

53
Q

How do u do gene therapy

A

altering the defective genes (mutated alleles)

54
Q

How do u treat a mutated gene caused by 2 recessive alleles

A

Add a dominant allele

This supplements the faulty alleles

55
Q

How do you treat a dominant allele

A

Silence it by adding a bit of dna in the middle of the allele to make it non functional

56
Q

How is the recombinant dna inserted into the cells for gene therapy

A

Vectors eg. Viruses, bacteria, liposomes

57
Q

What is somatic therapy

A

Altering the body cells (usually the ones most effected by the disease)
Doesn’t effect sex cells
Offspring could still inherit the fault alleles

58
Q

What is germ line therapy

A

Altering the sex cells

Every cell of every offspring produced would not inherit the disease as they would inherit the recombinant dna

59
Q

What is the ethical issue of gene therapy

A

People could use it for cosmetic purposes in offspring and reverse aging

60
Q

What is the purpose of gene probes

A

Locate specific alleles on chromosomes eg. Mutated alleles that cause a genetic disease

61
Q

What are dna probes

A

Short strands of dna
That has a specific base sequence that’s complementary to the base sequence on the target allele
So it will bind to the target allele
A label is attached so that it can detected

62
Q

Give 2 examples of labels on dna probes

A
Radioactive = detected using x ray film 
Fluorescence = uv light
63
Q

Describe how dna probes are used using electrophoresis

A
  1. Sample of dna digested into fragments using restriction endonuclease
  2. Fragments separated using electrophoresis
  3. Fragments transferred to a nylon membrane
  4. Incubated with a fluorescent label
  5. Allele present = probe will hybridise it
  6. Membrane exposed to uv
  7. Allele will be a Fluorescent band
64
Q

Describe how dna probes are used using a DNA microarray

A
  1. Sample of fluorescently labelled dna washed over array
  2. If the labelled dna contains the allele it will bind to the probe and stick to the array
  3. array is washed to remove unbound labelled dna
  4. Uv exposed
  5. Labelled dna will glow as a band
65
Q

What is a DNA microarray

A

Glass slide with microscopic spots containing different dna probes to different alleles

66
Q

What is the benefit of using DNA microarray

A

It screens lots of genes at once

67
Q

How is a dna probe produced

A

Need to sequence the allele you want

Use pcr to make lots of complementary parts to an allele

68
Q

What is the use of dna probes

A

Identify genetic conditions
Identify risk factors
Help decide which medicine would be most effective

69
Q

What is genetic counselling

A

Advising patients about risks, screening, results, most effective treatments

70
Q

How are personalised médecines used

A

Genes determine how you respond to certain drugs
Different people respons to the same drug differently
Médecines that are tailored to the dna which is the most effective

71
Q

What parts of the genome don’t code for proteins

A

Variable number tandem repeats

72
Q

What are variable number tandem repeats

A

Don’t code for amino acids

Repeat next to each other

73
Q

What makes vntr unique to each person

A

Repeated a different amount for different people a

Repeated in different places in the genome

74
Q

What is genetic fingerprinting

A

The number of times a sequence is repeated at different places in the genome compared between individuals

75
Q

Why is the probability of 2 genetic fingerprints being identical

A

The change of a person have the same number of vntr at the same places in the dna is low

76
Q

What is the function of electrophoresis

A

Sepeerate dna fragments

77
Q

What’s the method for dna electrophoresis

A
  1. Dna mixture in well in a slab of gel
  2. Covered in buffer solution that conducts electricity
  3. Electrical current passes through gel
  4. Dna is -ve charged so moves to the positive electrode
  5. Small dna moves faster than longer fragments so small moves further
78
Q

How do u carry out genetic fingerprinting

A
  1. Sample of dna from blood/saliva
  2. Pcr makes many copies of the DNA fragments that contain vntr
  3. Primers bind to either side of the repeats so the whole repeat is amplified
  4. Le both of nucleotides corresponds to how many repeats there are
  5. Fluorescent tags added to dna fragments
  6. Undergoes electrophoresis
  7. Exposed to uv light
  8. Bands shows as fluorescent bands
79
Q

How do u make sure the whole repeat is amplified

A

Add primers to either side of the repeat

80
Q

What does the length in nucleotides correspond to in dna fingerprinting

A

Corresponds to the number of repeats

81
Q

What does it mean if 2 fingerprints have a band in the same place

A

Same number of vntr at that place

82
Q

Give 5 reasons genetic fingerprinting is used for

A
  1. Determine genetic relationships eg. Paternity tests
  2. Determining genetic variability in a pop = greater the number that don’t match = the more diverse a pop is
  3. Forensic science
  4. Medical diagnosis
  5. Percent inbreeding of animals = find the least closely related organisms and breed together
83
Q

Why is genetic fingerprinting used in medical diagnosis

A

Used for when the specific allele mutation isn’t known because identifies a larger genetic pattern