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Manipulating Genomes 2 Flashcards

1
Q

Human genome

A

All of genetic material one contains that is dna in nucleus and mitochondria

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2
Q

Dna that codes for proteins

A

20-25000 regions of dna code for proteins
Only 2% - extrons

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3
Q

Introns

A

Large non coding regions of DNA
Removed from mRNA before translation

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4
Q

Satellite dna

A

Short sequences of dna that are repeated many times

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5
Q

Minisatellites

A

Sequence of 20-50 base pairs will be repeated from 50 to several hundred times
Occur at more than 1000 locations in genome
Known as variable number tandem repeats

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6
Q

Micro satellites

A

2-4 bases repeated only 5-15 times
Known as short tandem repeats

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7
Q

Patterns of dna satellites

A

Appear in same positions on chromosomes however the number of repeats vary (inherited mix)
Closer relation more similar the patterns

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8
Q

Producing dna profile

A

Extracting dna
Digesting the sample
Hybridisation
Seeing evidence

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9
Q

Extracting dna

A

Dna extracted from a tissue sample and use pcr in order to produce enough dna to develop a profile

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10
Q

Digesting the sample

A

Strands of dna cut into fragments using restriction endonucleases

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11
Q

Separating the dna fragments

A

Cut fragments of dna separated to form a clear pattern using electrophoresis

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12
Q

Hybridisation

A

Fluorescent dna probes are added in excess to fragments which are complementary to known dna sequence
Probes identify the micro satellites which are more varied than the minisattelites

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13
Q

Seeing the evidence

A

Sample placed under uv light so fluorescence glows - gives a pattern of bars which is unique to every individual apart from mz twins -THE DNA PROFILE

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14
Q

Restriction endonucleases

A

Enzymes which split dna into small fragments at specific sequences known as restriction site( complements active site)
Cut both dna strand
Allows scientist ability to cut at known and defined points in introns

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15
Q

Electrophoresis

A

Dna fragments put into well in gel strips ( one well contains known dna fragments)
Current passed through fragments move towards cathode due to -ve phosphate groups in dna - smaller frag closer to anode
Placed in alkali to denature

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16
Q

Southern blotting

A

Strands of dna transferred to nitrocellulose paper
Identical positions to the gel (alkali removed using absorbent paper)
Then hybridised

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17
Q

Pcr

A

Form of artificial dna replication
Semi conservative multiple complies
Amplifies sample

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18
Q

Pcr components

A

Pcr vial contains dna sample nucleotides 2x primers Taq DNA polymerase
Denature dna - heat 95 give single strands
Primers - cool mixture to 55 allows primers to anneal to dna
Synthesis - at 72 taq DNA polymerase strat synthesising new dna comp to og strand next to primer

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19
Q

Uses of dna profiling

A

Forensic science - evidence at scene
Paternity of a child
Identifying those at risk of particular disease

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20
Q

Dna sequencing

A

Process of determining the precise order of nucleotides within a dna molecule

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21
Q

Whyis dna sequencing used

A

Identifying genetic disorders - base sequence mutations
Identifying species using small conserved sections of DNA
Identifying evolutionary relationships
Pathogen analysis to develop vaccines
Proteonomics

22
Q

Dna barcoding ( identify species)

A

Use short section of DNA from conserved region of genome
648 base sequence of Mito. Dna which codes for enzyme analysed
Small enough to be quick varies enough to give clear differences

23
Q

Proteonomics

A

Study amino acids
Complexity of genotype phenotype relationship
Enables to predict the sequence of amino acids in all proteins it produces

24
Q

Chain termination technique

A

A method to find a base sequence of unknown genes using terminator bases

25
Terminator bases
Dideoxyribonucleotides - 1 less oxygen the deoxyribose Form comp pairs with deoxyribose bases Given fluorescenct tags to identify Dna polymerase stops replicating when it encounters the terminator bases
26
Dna sequencing technique
Pcr - single strand dna Dna polymerase builds up new strands of terminator base binds polymerase stops Results in different fragments stopped at every base Separated using capillary sequencing depending on chain length Fluorescent markers identify final base on each fragment Computer reassembles the genome
27
Synthetic biology
Design and construction of novel artificial biological pathways organisms or the redesign of existing natural biological systems
28
Synthetic bio techniques
Genetic engineering Use of biological systems in industry - creating enzymes Synthesis of new genes to replace faulty Synthesis of entire new organism
29
Gene therapy
Treatment of a disease by manipulating gene in a persons genetics
30
Germ line therapy
Changing genes in gametes to create off spring where all cells carry genetic modification (passed on to offspring)
31
Somatic cell therapy
Only specific group of body cells genetically modified/ different No modified genes are in gametes not passed onto offspring
32
Evaluation of somatic cell
Adv=uccessful treatments of leukaemia for example Dis=somatic cell has limited life replaced by stem cells that will have faulty allele
33
Germ line evaluation
Adv = successfully done w animals Dis= ethical concern desirable offspring illegal
34
Next generation sequencing
Gel plates and electrophoreses replaced by plastic plate called flow cell with dna fragments attached Produces stacks of DNA via chain termination Quicker cheaper and more efficient
35
GM crops
Developed to benefit grower and retailer Seeds need to be brought in each yr remain sterile - disadvantage poor Some modified to benefit poor farmers eg scuba rice resistant to floods
36
Pharming
Production of human medicine Mice removed genes develop cancer can study Bacteria making insulin Gene to cow egg protein harvested in milk
37
Ethical issues
Should animals be genetically engineered to act as models of human disease Is it right to put human genes in animals Religious beliefs Disease transfer from animal to human etc
38
Scid
Severe combined immunodeficiency disease Caused by faulty allele coding for enzyme ADA Essential for healthy immune system
39
Gene therapy for SCID
Removal of T cells and insertion of correct allele via vector Cells take up and clone allele Alternative injections of ADA May develop leukaemia
40
Cystic fibrosis
Thick mucus Caused by recessive allele that codes for CFTR Protein No channel proteins
41
Gene therapy for cystic fibrosis
Normal allele inserted into liposomes Lipid soluble can move through cell membrane of respiratory pathway Only last a week - somatic
42
Genetic engineering
Use of technology to change genetic material of one organism by taking genetic material from another organism
43
Isolating the gene gm
Use of restriction endonuclease to break at specific sites cutting dna unevenly leaving sticky ends This makes it easier to insert gene Or use reverse transcriptase to produce strand of comp DNA
44
Advantage of using reverse transcriptase
Makes it easier to identify desired gene due to particular cells making v specific types of mRNA
45
Formation of recombinant dna
Isolated dna inserted into bacterial plasmids Once plasmid gets into host cell combine w DNA to form recombinant dna Plasmids use as vectors contain marker gene eg antibiotic res which helps scientist determine whether taken up
46
Dna and plasmid fusion
Plasmid is cut open w same restriction endonuclease resulting in compl sticky ends Dna ligase forms phosphodiester bonds between sugar and phosphate groups on the dna strands Given second marker gene where dna fragment is placed so secondary marker gene doesn’t work
47
Transferring vector
Calcium rich sol and high temp - causes membrane to become more permeable Electroporation- current applied to bacteria make membrane v porous
48
Alternative gm techniques
Electrofusion- fusing of cells using electric current forming a hybrid polyploid cell (don’t survive in living animal) Agrobacterium tumefacians - plant produce a callous which can be grown into a new transgenic plant
49
Genetic engineering uses
Bacterial engineering to form medicines eg hormones antibiotics clotting factors etc In animals to try and cure human genetic diseases eg huntingtons or cystic fibrosis
50
Benefits and risks of genetic engineering
Microorg adv=make drugs dis= antibiotic resistance can spread Plant adv= high crop yield dis= damage environment Animal adv=improve understanding of genetic dis dis= welfare issues Humans adv=treat disorders dis= illegal and unknown risk

Biology (24 decks)

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