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Gene Tech Flashcards

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
What is the purpose of protecting groups
Stop unwanted branching | Make sure the nucleotides join at the right points
26
What does in Vivo mean
inside a living org
27
What does amplifying dna mean
Make lots of copies
28
Describe the method of amplifying dna in Vivo
1. Dna fragment inserted into a vector 1. Vector dna cut open using the same restriction endonuclease that was used to isolate the dna 2. Forms complementary sticky ends to the dna fragment 3. Vector dna and fragments are mixed with dna ligase 4. Vector dna and dna fragment join together = recombinant dna 5. Host cells take up the gene (become transformed) 6. Marker genes are inserted into the vector at the same time the dna fragment is added 7. Host cells grow on an agar plate. Any colony formed from a transformed cell will all have the recombinant dna and marker gene 8. Identify transformed cells eg. Uv light
29
What is ligase used for
Joins sticky ends of dna fragment to sticky ends of vector dna =ligation
30
How do host cell cells take up recombinant dna from a plasmid vector
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
31
How is recombinant dna taken up by host cells if the vector is a bacteriophage
The bacteriophage will inject its dna into the host cell
32
How do u produce proteins from the dna fragment
Vector needs to have specific promotors and terminator regions
33
What are promoter regions
Tell rna polymerase when to start productions mRNA
34
What does in vitro mean
Outside of living organisms
35
What is the process used in vitro dna amplifying
Pcr | Polymerase chain reaction
36
Décor cuve the method of in vitro dna amplifying
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
What is a primer
Short piece of dna that complementary to the bases at the start of the dna fragment
38
How many strands are made from each cycle of pcr
Double
39
What does DNA polymerase do
Lines up free nucleotides for complementary bases
40
How do u get plants to produce a desirable protein
``` 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
How do u make w genetically modified animal
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
How are promoter regions specific
Only activated in specific cell types | Can be used to control the which cells the gene is expressed in
43
Why are promoter regions so specific
Protein harvested more easily | If it was produced in the wrong cell it would be damaging
44
How is gm used in agriculture
Higher yields = stop famine More nutrients = stop malnourishment Pest resistance = fewer pesticides needed Reduce the costs to farmers
45
How is gm used in industry
Use biological catalysts Produced in large quantities Less money
46
How is gym used in médecine
Make vaccines and drugs quickly and cheaply | Make more efficient médecines = human insulin instead of pigs
47
How is gm in agriculture bad
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
How is gm used in industry bad
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
How is gym used in médecine bad
Companies who own tech may limit its use = but it could be saving lives Could be used unethically = designer babies
50
What are the ethical issues of gm
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
Name 4 benefits to gm for humans
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
What is gene therapy used for
Treating human diseases
53
How do u do gene therapy
altering the defective genes (mutated alleles)
54
How do u treat a mutated gene caused by 2 recessive alleles
Add a dominant allele | This supplements the faulty alleles
55
How do you treat a dominant allele
Silence it by adding a bit of dna in the middle of the allele to make it non functional
56
How is the recombinant dna inserted into the cells for gene therapy
Vectors eg. Viruses, bacteria, liposomes
57
What is somatic therapy
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
What is germ line therapy
Altering the sex cells | Every cell of every offspring produced would not inherit the disease as they would inherit the recombinant dna
59
What is the ethical issue of gene therapy
People could use it for cosmetic purposes in offspring and reverse aging
60
What is the purpose of gene probes
Locate specific alleles on chromosomes eg. Mutated alleles that cause a genetic disease
61
What are dna probes
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
Give 2 examples of labels on dna probes
``` Radioactive = detected using x ray film Fluorescence = uv light ```
63
Describe how dna probes are used using electrophoresis
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
Describe how dna probes are used using a DNA microarray
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
What is a DNA microarray
Glass slide with microscopic spots containing different dna probes to different alleles
66
What is the benefit of using DNA microarray
It screens lots of genes at once
67
How is a dna probe produced
Need to sequence the allele you want | Use pcr to make lots of complementary parts to an allele
68
What is the use of dna probes
Identify genetic conditions Identify risk factors Help decide which medicine would be most effective
69
What is genetic counselling
Advising patients about risks, screening, results, most effective treatments
70
How are personalised médecines used
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
What parts of the genome don’t code for proteins
Variable number tandem repeats
72
What are variable number tandem repeats
Don’t code for amino acids | Repeat next to each other
73
What makes vntr unique to each person
Repeated a different amount for different people a | Repeated in different places in the genome
74
What is genetic fingerprinting
The number of times a sequence is repeated at different places in the genome compared between individuals
75
Why is the probability of 2 genetic fingerprints being identical
The change of a person have the same number of vntr at the same places in the dna is low
76
What is the function of electrophoresis
Sepeerate dna fragments
77
What’s the method for dna electrophoresis
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
How do u carry out genetic fingerprinting
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
How do u make sure the whole repeat is amplified
Add primers to either side of the repeat
80
What does the length in nucleotides correspond to in dna fingerprinting
Corresponds to the number of repeats
81
What does it mean if 2 fingerprints have a band in the same place
Same number of vntr at that place
82
Give 5 reasons genetic fingerprinting is used for
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
Why is genetic fingerprinting used in medical diagnosis
Used for when the specific allele mutation isn’t known because identifies a larger genetic pattern

Biology 3 (10 decks)

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