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biology year 2 > Genome Projects and Gene Technologies > Flashcards

Genome Projects and Gene Technologies Flashcards

1
Q

define genome

A

complete set of genetic material that organism has

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2
Q

define proteome

A

complete set of proteins that organism can make

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3
Q

human genome project

A
  • sequenced whole genome
  • only sequence short fragments at once
  • split genome into small sections - sequence - put back in order
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4
Q

how are sequencing methods being constantly updated

A
  • chain termination method

to

  • pyrosequencing
  • automated
  • faster cheaper
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5
Q

predicting AA sequences in simple organisms

A
  • few regulatory genes
  • little non coding DNA
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6
Q

predicting AA sequences in complex organisms

A
  • non-coding dna e.g. VNTRs
  • regulatory gene turns on/off other genes
  • lots of non-coding dna
  • hard to determine proteins from dna
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7
Q

uses of genome projects

A
  • understand evolutionary relationships
  • medicine - understand antigens to develop new vaccines
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8
Q

define gene therapy

A

changing faulty alleles that cause genetic disease

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9
Q

gene therapy for disease caused by dominant allele

A
  • sufferes = hetero
  • alreadu have functional allele
  • silence dominant allele
  • use vector to add dna fragment into dominant allele
  • dominant allele not transcribed
  • recessive allele expressed
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10
Q

gene therapy for disease caused by recessive allele

A
  • sufferers = homo
  • use vector to add functional allele to DNA
  • dominant allele expressed
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11
Q

2 types of gene therapy

A
  • germ line
  • somatic
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12
Q

germ line gene therapy

A
  • changing alleles of gametes
  • all future offspring inherit
  • illegal
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13
Q

somatic gene therapy

A

changing allele of body cells

  • offspring dont inherit changes
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14
Q

problems with gene therapy

A
  • alleles inserted into wrong locus
  • could silence wrong gene by mistake
  • could cause cancer
  • gene over expressed
  • non-medical uses
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15
Q

3 ways to isolate target genes

A
  • restriction enzymes
  • reverse transcriptase
  • gene machine
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16
Q

restriction enzymes

A
  • RE cut dna at specific palindromic sites = restriction sites
  • if restriction site either end, RE used to cut it out
  • leaves dna with sticky ends (unpaired bases)
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17
Q

reverse transcriptase

A
  • cells have 2 copies of each gene - hard to access
  • enzyme that does transcription backwards

DNA + RNA polym –> mRNA

mRNA + reverse transcriptase —> cDNA

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18
Q

gene machine

A
  • join 25 nucleotides together at once
  • forms oligonucleotides
  • join oligonucleotides = synthetic gene
  • design own genes/ proteins
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19
Q

what is PCR used for?

A
  • amplify DNA (make millions of copies)

known as IN VITRO dna amplification

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20
Q

PCR ingredients

A
  • DNA sample
  • free DNA nucleotides
  • primers
  • DNA polymerase (taq polymerase)
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21
Q

what are primers?

A

short sequences of DNA that are comp to start of dna sample

  • used to select which part of dna copied
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22
Q

PCR method

A
  • heat to 95 degrees - break H bonds & make DNA single stranded
  • cool to 50 degrees - allow primers bind - comp base pairing - dna double stranded - dna polymerase can bind
  • heat to 70 degrees - dna polymerase adds comp nucleotides- forms phosphodiester bonds
  • repeat - each cycle doubles DNA

1-2-4-8-16

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23
Q

what is a vector

A

something thats used to move DNA from one place to another

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24
Q

what are virus

A

bacteriophage

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25
Q

what are plasmids

A

double stranded loop of DNA that transfers genes between bacteria

26
Q

how do you insert a gene into a vector

A
  • use same restriction enzyme to cut plasmid
  • sticky ends will be comp
  • dna ligase reforms phosphodiester bond
  • forms recombinant dna (dna from more than one source)
27
Q

how do you insert vector into bacteria

A
  • ice cold CaCl2
  • heat shock

INC PERM OF BACTERIAL CELL WALL

28
Q

what is a transgenic organism

A

contains rDNA

29
Q

what is electrophoresis used for

A

using electricity to separate DNA fragments by length

30
Q

electrophoresis

A
  • DNA negatively charged
  • attach fluorescent label or stain DNA frag
  • put DNA frag into well at negative end
  • turn on current
  • dna frag attracted to positive electrode
  • small frag faster (further)
31
Q

in electrophoresis how do you calibrate the scale

A

wiring pieces of dna of known length

32
Q

3 fragments =

A

2 cuts

33
Q

what are marker genes

A

genes paired with target genes to check if vector inserted properly

34
Q

give types of marker gene

A
  • UV fluorescence
  • antibiotic resistance
35
Q

what do transformed bacteria contain

A

rDNA (target gene and marker gene)

36
Q

only bacteria that have accepted the vector will..

A
  • will fluoresce under UV
  • will be able to survive in culture with antibiotic
37
Q

what do you do once transformed bacteria have been identified?

A

select and culture transformed bacteria

38
Q

what is a dna probe

A

a short sequence of dna with label attached that is complementary to a specific allele/mutation/gene

39
Q

what are dna probes used for

A

used to test if sample of dna contains specific sequence

40
Q

how dna probes work

A
  • attach label to dna probe
  • if sequence (disease causing allele) present
  • dna probe will hybridise and stick to dna sample with comp base
  • rinse to remove unhybridised dna probes
  • view labels (uv fluorescene, radioactive) - dna invisible
41
Q

what must dna be when using dna probes

A

single stranded

42
Q

dna microarray

A
  • label attached to human dna (not probe)
  • test multiple alleles/mutations at once
  • many dna probes attached to tile in grid (know positions)
  • attach patients dna with label
  • dna will hybridise to comp dna
  • rinse —> view labels
43
Q

uses in agriculture (soya beans, golden rice)

A
  • express protein from bacteria
  • protein is toxic to insects
  • fewer insects eat soya plant
  • gene from corn —> rice
  • express vitamin A
44
Q

advantages of transformed organisms in agriculture

A
  • use less chemical pesticide
  • more efficient food chain
  • prevents blindness caused by vitamin A deficiency
45
Q

disadvantages of using transformed organisms in agriculture

A
  • monoculture of crop has low genetic diversity
  • susceptible to enviro factors e.g. disease
  • have to buy seeds every year
  • dec in biodiversity
46
Q

uses of transformed organisms in industry and research

A
  • making enzymes
  • e.g. rennin, lipase
  • transformed pathogens to TREAT disease
  • attack other pathogens but dont infect humans
47
Q

advantages of using transformed organisms in industry and research

A
  • reduce energy and cost
  • fast and cheap production
  • treat disease
  • pathogens wont develop resistance
  • reduce suffering
48
Q

disadvantages of using transformed organisms in industry and research

A
  • could mutate and infect humans
  • could be used in war
49
Q

uses of transformed organsism in medicines (insulin)

A
  • transform bacteria to express proteins
  • mammals can also be transformed to produce useful prod in their milk
50
Q

advantages of using transformed organisms in medicine

A
  • makes human proteins
  • cheaper and easier than making proteins synthetically
51
Q

disadvantages of using transformed organisms in medicine

A
  • possible unexpected problems e.g. cancer in animals
  • using animals as commodities
52
Q

what is genetic counselling used for

A
  • identify carriers of genetic disease
  • specific allele
  • most effective treatment
53
Q

genetic counselling

(USES OF DNA PROBES)

A
  • healthcare professionals advise people/parents about risks
  • passing on inheritable diseases
  • developing diseases later in life
  • e.g. angelina jolie
  • BRCA1- 87% chance of developing breast cancer
  • double mastectomy
  • make informed decisions about treatments and preventions
54
Q

genetic screening

(USES OF DNA PROBES)

A
  • parents can see if they are carriers of recessive alleles (IVF)
  • can diagnose and treat before symptoms show
  • e.g. babies screened for CF
55
Q

personalised medicine

(USES OF DNA PROBES)

A
  • if doctor knows patients genotype - give best drugs for them
  • mutation X - drug X
  • mutation Z - drug Z
  • change drugs if know patient has allele that causes side effect with normal drug
56
Q

what is genetic fingerprinting

A

identifying individuals by comparing difference in Variable Number Tandem Repeats (VNTRs)

57
Q

what does non-coding dna contain lots of

A

VNTRs

  • dont code for proteins
  • dont affect phenotype - vary more than coding dna
  • number of repeats of each VNTR varies
58
Q

relatedness

USES OF GFP

A
  • more closely related two individuals - high percentage match of VNTRs
  • conservation of endangered species
  • avoid inbreeding by matching individuals who = least related
  • maintains genetic diversity
59
Q

forensic use

USES OF GFP

A
  • samples of dna from crime scene and suspects
  • amplify using PCR
  • run gel electrophoresis
  • chance of 2 individuals having same number of VNTRs very low
60
Q

medical diagnosis

USES OF GFP

A
  • test for specific combinations of alleles
  • used to diagnose disorders
    e. g. embryo screening for IVF

biology year 2 (29 decks)

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