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Biology > 6.1.3 Manipulating Genomes > Flashcards

6.1.3 Manipulating Genomes Flashcards

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1
Q

what is SD’s - PAGE Electrophoresis

A

method used to seperate proteins

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2
Q

what can electrophoresis be used for

A

to analyse mixes of proteins to determine what is present

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3
Q

What is DNA profiling

A

short tandem repeats are compared as people sharing an str is very low

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4
Q

What is a short tandem repeat

A

repetitive sequences of 10 -100 base pairs

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5
Q

Steps of DNA fingerprinting

A

Extract DNA
cut DNA using resitriction enzyme
seperate DNA fragments using electrophoresis
Then do the same with the individual being compared

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6
Q

How to do southern blotting

A

-cut DNA out of gel and purify it
- or place nitrocellulose sheet over gel and press down leave over night and the DNA transfers to the sheet

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7
Q

What is a probe in Southern Blotting

A

single stranded and complementary to DNA your trying to find

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8
Q

What does a probe do

A

bind to DNA its complementary to and lets out a fluoresent colour to identify it easily

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9
Q

How are probes labelled

A

radioactively or with fluorecent colour

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10
Q

DNA Sequencing definition

A

what order ATCG go

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11
Q

Applications of DNA Sequencing

A

identify mutations
identify genetic disorders
used to help create vaccines
identify common ancestors
prove paternity
find desired characteristics and genetic engineer them

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12
Q

Sanger sequencing method

A

4 test tubes
put fragmentated DNA to be sequenced, bases(A,T.C.G), DNA polymerase and primers into all the test tubes
Add modified bases (ddNTPs) which are radioactive- one for each testube

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13
Q

What happens during sanger sequencing

A

DNA polymerase adds bases onto primer and produces many new strands
Everytime a ddNTP is added it terminates the chain so fragment ends

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14
Q

High throughput sequencing

A

first automated DNA sequencing machine was made in 1986
used fluoresence rather than autoradiography

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15
Q

Bioinformtives

A

develops and uses computer software to analyse, store and organise biological data

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16
Q

What is the data in bioinformatives

A

univeral
shared internationallly
DNA, RNA, Protein

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17
Q

Genome

A

total complement of an organisms genes

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18
Q

what does Dna sequencing do

A

determines the order of nucleotides in a dna sample

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19
Q

common techniques of gene technologies

A

restriction endonucleous
PCR
Electrophoresis
Dna probes and microarrays
dna profiling
dna sequencing
genetic engineering
gene therapy

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20
Q

DNA Fragmentation method

A

Restriction endonucleases recognise palindromic sequences and cut the dna and these sites
leaves overhanging sticky ends
each enzyme is specific to a sequence and is complementary to their active site

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21
Q

what does PCR do

A

makes more copies of a section of DNA
amplifies the dna

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22
Q

Uses of PCR

A

identify viral rna/dna in patient
forensics
ancient organisms
epidermiology
detect and identify mutations e.g cancerous mutations

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23
Q

PCR mixture

A

sample of dna
excess of 2 primers
dna polymerase
4 dna bases
appropriate buffer

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24
Q

4 steps of PCR

A

denaturation
annealing
elongation
amplification

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25
Q

What happens in the 1st step of PCR

A

denaturation
DNA is heated at 95 degrees to break hydrogen bonds between bases to get a single strand

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26
Q

what happens in the 2nd step of PCR

A

annealing
cool to 68 degrees
primers will anneal to dna by complementary base pairing

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27
Q

what happens in 3rd step of PCR

A

elongation
heat to 72 degrees to allow dna polymerase to extend the primers and copies of the target dna are synthesised using target strands as templates

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28
Q

what happens in the 4th step of PCR

A

amplification
each time the above 3 steps are repeated the number of DNA strands doubles

29
Q

what does comparitive gene mapping do

A

identifies importance of difference genes
can see how tiny changes in gene causes different features
comparing species identifies evolutionary relationships

30
Q

synthetic biology

A

designs and builds biological devices and systems

31
Q

why is knowing the dna sequence good

A

can lead to modelling the amino acid sequence

32
Q

examples of synthetic biology uses

A

synthetic new proteins
biosensors
nanotechnology
storing information
new medicines

33
Q

genetic engineering definition

A

direct manipulation of an organisms genes

34
Q

what can genetic engineering involve

A

addition of genes
removal of genes
silencing of genes by blocking genes expression

35
Q

what is dna ligase used as

A

a sealing enzyme

36
Q

what is a dna probe

A

short single strand dna fragment
labelled with 32P or fluoresence
used to locate a gene

37
Q

Method to genetically engineering an insulin gene

A

Cut out insulin gene using restriction enzyme and leave sticky ends
cut plasmid from bacteria with the same restriction enzyme
insert gene into plasmid using DNA ligase
transgenic bacteria reproduce in a fermenter
genetically engineered insulin is then produced by the bacteria

38
Q

Why do you use the same restriction enzyme to cut the plasmid and the insulin gene

A

so they have complementary sticky ends that overhang

39
Q

what enzyme is used to insert insulin gene into the plasmid

A

DNA ligase

40
Q

Stages of genetic engineering

A

obtain gene of interest
insert gene into vector
insert vector into cell
or directly insert gene into the cell

41
Q

How do you obtain the gene of interest if you don’t know the sequence of gene

A

extract mRNA
use reverse transcriptase to make single stranded cDNA
then use DNA polymerase to make it double stranded

42
Q

How do you obtain the gene of interest if you know the sequence of the gene

A

can be produced using automated synthesiser
amplify gene from genomic DNA using PCR
can cut it out of genomic DNA using restriction enzyme

43
Q

Types of vectors you can insert a gene into

A

seal in a virus
use a bacterial plasmid using same method as insulin

44
Q

ways to insert the vector into a cell

A

heat/cold shock
electroporation
electrofusion
transfection
T1 plasmid

45
Q

Differences between somatic and germ line gene therapy

A

Somatic is body cells Germ line is gametes
Somatic cures you germ line cures your offspring
Somatic causes specific cells to have gene germ line all cells have the gene

46
Q

How does inserting a new gene into a chromosome could affect the functioning of other genes in that chromosome

A

Gene expression changes
Newly genes switched on or off
New gene could disable functioning gene if its inserted

47
Q

GM crop characteristics

A

Good growth conditions
Pathogen resistance
Pesticide resistance
Herbicide resistance
Nutritional value and good appearance
Long shelf life

48
Q

Pros and cons of good growth conditions for GM crops

A

High yield
Expensive

49
Q

Pros and cons of pathogen resistance for GM crop

A

High yield
Transfer of gene to wild species

50
Q

Pros and cons pesticide resistance of GM crops

A

High yield helps poor farmers
Biaccumalate pests can become resistant

51
Q

Pros and cons of herbicides resistance for GM crops

A

High yield
Create superweeds which are resistant to herbicides

52
Q

Pros and cons of nutritional value and appearance for GM crops

A

More likely to be brought
Good for human health
Medicine development
May be allergenic for some people

53
Q

Pros and cons of long shelf life for GM crops

A

Easy to import and export
Better customer satisfaction
Commercial value decreases as people don’t need to buy them as often

54
Q

Gene therapy

A

Involves treating genetic diseases by altering a patients natural genotype

55
Q

How to do gene therapy for somatic cell therapy

A

Normal allele is incorporated into a loop of dna which is attached to liposomes
Liposome-DNA complex is sprayed as an aerosol of fine droplets into nose of cystic fibrosis sufferers

56
Q

Advantage of somatic gene therapy

A

Better quality of life
Removes symptoms

57
Q

Disadvantages of somatic gene therapy

A

Not a cure
Has to be done on a regular basis
Pass on abnormal gene to offspring

58
Q

Advantage of germ line gene therapy

A

Cure for children / offspring

59
Q

Disadvantages of germ line gene therapy

A

Not a cure for self
Expensive as it’s ivf

60
Q

Pharming

A

Using GM animals to produce pharmaceutical

61
Q

What happens in pharming

A

Add or remove genes so the animal developers certain discuss to act qs models in development of new treatments

62
Q

Example of pharming

A

Knock out mice have genes deleted so they develop cancer

63
Q

Action of a restriction enzyme

A

Cuts the dna into fragments at a certain recognition site leaving sticky ends

64
Q

2 ways in which bacteria which take up modified plasmid can be identified

A

fluorescent substance and which ever plasmid is fluorecent contain it
antibiotic resistance gene introduced and survivors have plasmid

65
Q

restriction enzyme

A

cuts dna at specific site leaving sticky ends

66
Q

characteristics of restriction enzymes target site

A

palindromic
pairs
specific sequence

67
Q

advantages of obtaining human insulin by genetic engineering

A

faster production
less chance of it getting rejected
easier to grow on mass scale

68
Q

3 reasons researchers may be more concerned about potential risk of dna vaccines compared with protein based vaccines

A

could trigger autoimmune response
could undergo mutations
create new disease

69
Q

what is recombinant dna

A

dna from 2 differnt dources joined

Biology (25 decks)

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