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Biology > 6.1.3 Manipulating Genomes > Flashcards

6.1.3 Manipulating Genomes Flashcards

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
What happens in the 1st step of PCR
denaturation DNA is heated at 95 degrees to break hydrogen bonds between bases to get a single strand
26
what happens in the 2nd step of PCR
annealing cool to 68 degrees primers will anneal to dna by complementary base pairing
27
what happens in 3rd step of PCR
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
28
what happens in the 4th step of PCR
amplification each time the above 3 steps are repeated the number of DNA strands doubles
29
what does comparitive gene mapping do
identifies importance of difference genes can see how tiny changes in gene causes different features comparing species identifies evolutionary relationships
30
synthetic biology
designs and builds biological devices and systems
31
why is knowing the dna sequence good
can lead to modelling the amino acid sequence
32
examples of synthetic biology uses
synthetic new proteins biosensors nanotechnology storing information new medicines
33
genetic engineering definition
direct manipulation of an organisms genes
34
what can genetic engineering involve
addition of genes removal of genes silencing of genes by blocking genes expression
35
what is dna ligase used as
a sealing enzyme
36
what is a dna probe
short single strand dna fragment labelled with 32P or fluoresence used to locate a gene
37
Method to genetically engineering an insulin gene
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
Why do you use the same restriction enzyme to cut the plasmid and the insulin gene
so they have complementary sticky ends that overhang
39
what enzyme is used to insert insulin gene into the plasmid
DNA ligase
40
Stages of genetic engineering
obtain gene of interest insert gene into vector insert vector into cell or directly insert gene into the cell
41
How do you obtain the gene of interest if you don't know the sequence of gene
extract mRNA use reverse transcriptase to make single stranded cDNA then use DNA polymerase to make it double stranded
42
How do you obtain the gene of interest if you know the sequence of the gene
can be produced using automated synthesiser amplify gene from genomic DNA using PCR can cut it out of genomic DNA using restriction enzyme
43
Types of vectors you can insert a gene into
seal in a virus use a bacterial plasmid using same method as insulin
44
ways to insert the vector into a cell
heat/cold shock electroporation electrofusion transfection T1 plasmid
45
Differences between somatic and germ line gene therapy
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
How does inserting a new gene into a chromosome could affect the functioning of other genes in that chromosome
Gene expression changes Newly genes switched on or off New gene could disable functioning gene if its inserted
47
GM crop characteristics
Good growth conditions Pathogen resistance Pesticide resistance Herbicide resistance Nutritional value and good appearance Long shelf life
48
Pros and cons of good growth conditions for GM crops
High yield Expensive
49
Pros and cons of pathogen resistance for GM crop
High yield Transfer of gene to wild species
50
Pros and cons pesticide resistance of GM crops
High yield helps poor farmers Biaccumalate pests can become resistant
51
Pros and cons of herbicides resistance for GM crops
High yield Create superweeds which are resistant to herbicides
52
Pros and cons of nutritional value and appearance for GM crops
More likely to be brought Good for human health Medicine development May be allergenic for some people
53
Pros and cons of long shelf life for GM crops
Easy to import and export Better customer satisfaction Commercial value decreases as people don't need to buy them as often
54
Gene therapy
Involves treating genetic diseases by altering a patients natural genotype
55
How to do gene therapy for somatic cell therapy
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
Advantage of somatic gene therapy
Better quality of life Removes symptoms
57
Disadvantages of somatic gene therapy
Not a cure Has to be done on a regular basis Pass on abnormal gene to offspring
58
Advantage of germ line gene therapy
Cure for children / offspring
59
Disadvantages of germ line gene therapy
Not a cure for self Expensive as it's ivf
60
Pharming
Using GM animals to produce pharmaceutical
61
What happens in pharming
Add or remove genes so the animal developers certain discuss to act qs models in development of new treatments
62
Example of pharming
Knock out mice have genes deleted so they develop cancer
63
Action of a restriction enzyme
Cuts the dna into fragments at a certain recognition site leaving sticky ends
64
2 ways in which bacteria which take up modified plasmid can be identified
fluorescent substance and which ever plasmid is fluorecent contain it antibiotic resistance gene introduced and survivors have plasmid
65
restriction enzyme
cuts dna at specific site leaving sticky ends
66
characteristics of restriction enzymes target site
palindromic pairs specific sequence
67
advantages of obtaining human insulin by genetic engineering
faster production less chance of it getting rejected easier to grow on mass scale
68
3 reasons researchers may be more concerned about potential risk of dna vaccines compared with protein based vaccines
could trigger autoimmune response could undergo mutations create new disease
69
what is recombinant dna
dna from 2 differnt dources joined

Biology (25 decks)

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