Topic Eight - DNA Technology Flashcards

1
Q

what effect does deletion have on DNA?

A

frame shift to left

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2
Q

what effect does addition have on DNA?

A

frame shift to right

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3
Q

what effect does duplication have on DNA?

A

frame shift to right

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4
Q

what effect does inversion have on DNA?

A

makes a different/ non-functional protein

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5
Q

what effect does substitution have on DNA?

A
  • makes a stop codeon = protein cut short
  • codes for a different a.a = different/non-functional protein
  • no effect (degenerate code)
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6
Q

what does translocation do to the DNA?

A

cuts one chromosome short and adds the gene onto another chromosome

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7
Q

what effect does translocation have on DNA?

A

changes gene expression

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8
Q

what cell types can totipotent stem cells differentiate into?

A

any cell type

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9
Q

what cell types can pluripotent stem cells differentiate into?

A

many cell types

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10
Q

what cell types can multipotent stem cells differentiate into?

A

limited cell types

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11
Q

what cell types can unipotent stem cells differentiate into?

A

1 cell type

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12
Q

where are pluripotent cells found, and what is the ethical issue?

A

embryo - destroyed
foetus - aborted/miscarried

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13
Q

where are multipotent cells found, and what is the ethical issue?

A

adult bone marrow - painful + risky
umbilical cord - naturally detaches

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14
Q

what are iPS cells?

A

unipotent cells that are genetically altered to have characteristics of embryonic cells (pluripotent)

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15
Q

what does an active binding site on a transcription factor do?

A

stimulates transcription

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16
Q

what does an inactive binding site on a transcription factor do?

A

inhibits transcription

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17
Q

how does oestrogen initate transcription?

A
  • binds to receptor on transcription factor
  • TF’s tertiary structure changes = complementary to DNA
  • Tf moves to nucleus
  • binds to a specific DNA sequence
  • imitates transcription
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18
Q

how is oestrogen linked to breast cancer?

A
  • after menopause more oestrogen produced
  • more transcription
  • tumour
  • increase in oestrogen conc.
  • increases tumour development
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19
Q

definition: epigenetics

A

heritable changes in gene function without changes to the base sequence of DNA

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20
Q

what does the epigenome do?

A

determines the shape of the DNA-histone complex

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21
Q

what is the epigenome?

A

the layer of tags that covers DNA + histones

22
Q

what can effect the epigenome?

A

stress, diet, exercise

23
Q

what happens when the DNA is tightly packed?

A

Tf + RNA polymerase can’t reach genes
= transcription is inhibited = gene is inactive

24
Q

what happens when the DNA is lossley packed?

A

Tf + RNA polymerase can reach genes
= transcription is stimulated = gene is active

25
where are tags found on the DNA?
methyl groups - bind to DNA acetyl groups - bind to histones
26
what is acetylation?
addition of an acetyl molecule that is donated from Acetyl CoA
27
what is methylation?
addition of CH3 its added to the C base of DNA
28
what causes translation to be inhibited?
increased methylation decreased acetlyation
29
what causes translation to be stimulated?
decreased methylation increased acetylation
30
how does gestational diabetes occur?
- fetus is exposed to high conc. of glucose - = epigenetic changes in daughter's DNA - increases chance she gets it
31
what are the two types of RNAi, and their diffrences?
miRNA (single stranded) SiRNA (double stranded)
32
how does RNAi stop translation?
miRNA / 1 strand of SiRNA, binds w a protein complex = RNA induced silencing complex (RISC) RISC binds with mRNA, stoping translation by: breaking mRNA OR prevents ribosomes from attaching to mRNA
33
what are the diffrences between benign and malignant tumours?
benign: - localised effect - less life threatening - removed by surgery - rarely reoccur malignant: - systemic effect - more life threatening - removed by surgery and treatment - more likely to reoccur
34
what do tumour suppressor genes do?
- slow/prevent cell division - repair DNA mistakes - apoptosis
35
what happens when a tumour suppressor gene is inactive?
HUGE increase in cell division = tumour
36
how do tumour suppressor genes become inactive?
- gene mutation - increased methylation
37
what do proto-oncogenes do?
increase cell division
38
what are oncogenes?
mutated proto-oncogenes
39
what do oncogenes do?
HUGE increase in cell division = tumour
40
what is gel electrophoresis used for?
to seperate DNA - by mass RNA - by mass proteins - by mass or charge
41
what are the three ways a protein fragment is produced?
restriction endonuclease reverse transcriptase gene machine
42
what are the three ways a DNA fragment is produced?
restriction endonuclease reverse transcriptase gene machine
43
how do restriction endonuclease create DNA fragments?
they cut DNA @ recognition sites = blunt ends or sticky ends
44
how does reverse transcriptase create DNA fragements?
- adds nucleotides to mRNA molecule = new strand called cDNA - enzyme destroys mRNA strands - 2nd strand is built by DNA polymerase = complete DNA fragment
45
how does the gene machine create DNA fragements?
- sequence is generated from the database - safety check - computer designs oligonucleotides - oligonucleotides join to make a gene
46
what happens in in vivo cloning?
- DNA fragment is prepared - insert fragment into the vector, using DNA ligase - place into host cell (use Ca2+ & heat shock) - identify bacteria with recombinant DNA (enzyme, fluorescence, antibiotic resistance) - multiply bacteria
47
what is replica plating?
- taking sample of bacteria into a new plate with the antibiotic, bacteria is at the same place - bacteria that dies = has the recombinant DNA
48
what is the process in in vitro cloning?
PCR 1) heated to 95°C -> breaks all the H bonds 2) cooled + DNA primers attach 3) heated, taq polymerase + nucleotides attach to each strand
49
what are the two ways of labeling DNA probes?
radioactive and fluorescent
50
how do you screen for genetic disorders?
1. multiply the DNA probe (PCR) 2. heat the DNA sample 3. Add probes + cool mixture 4. wash DNA sample 5. check for attachment
51
what are VNTRs?
repetitive, non-coding DNA sequences that are directly next to each other
52
how is genetic fingerprinting done?
1. collect DNA sample 2. use PCR to multiply DNA 3. restriction enzymes are used to produce VNTR DNA fragments 4. use gel electrophoresis to separate VNTRs 5. add alkali to separate VNTRs into single strands 6. add complementary DNA probes to identify VNTRs 7. Compare VNTR patterns