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Biology Y13 > Gene technology > Flashcards

Gene technology Flashcards

1
Q

Gene definition

A

DNA base sequence which codes for either the amino acid sequence of a polypeptide or a functional RNA

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2
Q

Genome definition

A

All the DNA in a cell

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3
Q

Proteome definition

A

The range of proteins a cell can make

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4
Q

Intron definition

A

Non-coding DNA WITHIN a gene

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5
Q

Non-coding multiple repeats definition

A

Non-coding DNA BETWEEN genes

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6
Q

What kind of DNA is the DNA of chloroplasts and mitochondria the same as?

A

Prokaryotic

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7
Q

What do genome projects determine?

A

The DNA base sequence of an organism’s genome

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8
Q

How can genome projects be used to determine an organism’s proteome?

A

Allow the amino acid sequence of a protein to be determined

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9
Q

Why is it easier to determine the proteome of prokaryotic DNA?

A

No introns, shorter so faster

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10
Q

How can genome projects be used in medical research?

A

Can identify the antigens of microorganisms for use in vaccine production

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11
Q

What 2 features in eukaryotes makes it difficult to translate the genome into the proteome?

A

Non-coding DNA and regulatory genes

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12
Q

How is a DNA base sequence determined?

A

Chain termination sequencing

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13
Q

What 4 components are needed in chain termination sequencing?

A
  1. Sample of DNA
  2. 4 normal DNA nucleotides
  3. DNA polymerase
  4. Primer
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14
Q

What is added in chain termination sequencing that fluoresce a different colour?

A

Chain terminating nucleotides which can’t form a phosphodiester bond and so stop DNA replication

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15
Q

What are DNA fragments run on in chain termination sequencing?

A

Electrophoresis gel

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16
Q

Recombinant DNA definition

A

DNA made from 2 or more different species

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17
Q

What are the 3 methods of producing a DNA fragment in in vivo gene cloning?

A
  1. Reverse transcriptase
  2. Restriction enzymes
  3. Gene machine
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18
Q

What does reverse transcriptase do and how does this method produce a DNA fragment (3 steps)?

A

Forms DNA from mRNA
1. mRNA coding for (insulin) removed (from beta cells in human pancreas)
2. Reverse transcriptase makes cDNA from mRNA
3. DNA polymerase makes cDNA double-stranded

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19
Q

How do restriction enzymes produce a DNA fragment?

A

Hydrolyse phosphodiester bonds at recognition sites which forms sticky ends

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20
Q

Why is the gene machine more efficient in producing a DNA fragment than the other 2 methods?

A
  1. Very accurate
  2. Faster as no need to isolate DNA/RNA
  3. No introns so can be expressed by prokaryotes
  4. Any sequence of nucleotides can be made
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21
Q

What must be added to a DNA fragment before it is inserted?

A

Promoter and terminator

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22
Q

Vector definition

A

Carries the gene into the cell

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23
Q

How is a gene bound to a plasmid (3 steps)?

A
  1. Restriction enzymes cut open the plasmid
  2. Sticky ends of the plasmid are complementary to the sticky ends of the gene
  3. DNA ligase joins nucleotides of plasmid and inserted gene
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24
Q

How is a DNA fragment introduced to a host cell?

A

Heat shock makes the cell’s membrane more porous so the plasmid/gene can pass through more easily

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25
Q

What does a marker gene do?

A

Identifies which cells have taken up the plasmid/gene

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26
Q

How does an antibiotic resistance marker gene identify bacteria containing the plasmid?

A

Only cells containing the plasmid survive

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27
Q

Which process identifies which plasmids contain the recombinant DNA?

A

Replica plating (ampicillin, tetracycline, marker gene disruption etc.)

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28
Q

Why is PCR used?

A

Amplifies a particular DNA base sequence

29
Q

What are the 3 differences between PCR and DNA replication?

A
  1. Heat is used to break hydrogen bonds instead of DNA helicase
  2. Taq polymerase is used instead of DNA polymerase
  3. Primers are used to replicate a section of DNA instead of whole molecule
30
Q

Why is the tube containing ingredients kept on ice in PCR?

A

Reduces enzyme activity to prevent damage to DNA

31
Q

What 4 ingredients are needed in PCR?

A
  1. DNA we want to copy
  2. DNA nucleotides
  3. Taq DNA polymerase
  4. Primers
32
Q

Primer definition

A

Short, single-stranded DNA with a complementary base sequence to the start and end of the gene to be copied

33
Q

Why are primers needed in PCR?

A

Mark where DNA replication should occur from, allow DNA polymerase to bind, keep the strands separated

34
Q

Why are 2 different primers needed in PCR?

A

DNA has 2 strands which different complementary base sequences

35
Q
  1. Why is the sample of DNA first heated to 95°C in PCR?
A

Breaks the hydrogen bonds to separate strands

36
Q
  1. Why is the sample of DNA then heated to 55°C in PCR?
A

Allows primers to bind

37
Q
  1. Why is the sample of DNA then reheated to 72°C in PCR?
A

Taq DNA polymerase binds and joins DNA nucleotides to create a new strand

38
Q

How is the number of DNA molecules made in each cycle calculated in PCR?

A

2^x

39
Q

Why does DNA replication eventually stop in PCR?

A

All of the primers or DNA nucleotides were used up

40
Q

What is one use of PCR?

A

Used to make many copies of DNA found at a crime scene for genetic fingerprinting

41
Q

What is gene therapy used for?

A

Used to cure a genetic disease caused by a mutated gene

42
Q

How does gene therapy cure a genetic disease?

A

Functional gene is inserted into DNA of a cell which expresses the mutated gene so functional protein is produced

43
Q

What are the 3 ways of inserting a gene into a cell in gene therapy?

A
  1. Using a virus
  2. Liposomes (phospholipid vesicles containing DNA) fuse with cell surface membrane
  3. Recombinant plasmid injected into cell
44
Q

What is somatic gene therapy?

A

Gene is inserted into somatic cells (normal adult body cells) which express the mutated gene

45
Q

Which type of somatic cells would be best to target in gene therapy and why?

A

Adult stem cells so all daughter cells contain the functional gene and less repeat treatments are needed

46
Q

What is germ line gene therapy?

A

Gene inserted into sperm or egg cells

47
Q

What are the advantages and disadvantages of GERM LINE gene therapy?

A

Advantages: functional gene passed on to offspring so genetic disease isn’t inherited
Disadvantages: illegal as could be used for cosmetic uses, can cause side effects

48
Q

What are the 4 disadvantages of gene therapy?

A
  1. Multiple treatments required as cells die - short term
  2. Still some faulty cells present
  3. Viruses can cause immune responses/side effects
  4. DNA may insert into the middle of another gene, forming a non-functional protein
49
Q

Why is gel electrophoresis used?

A

Separates out DNA fragments by length and charge

50
Q

What makes DNA negatively charged?

A

Phosphate groups

51
Q

How does gel electrophoresis separate DNA fragments?

A

DNA attracted to positive electrode, shorter DNA fragments travel the furthest

52
Q

How are unknown lengths of DNA fragments determined using gel electrophoresis?

A

Compared to KNOWN lengths of DNA fragments on the gel

53
Q

How can PROTEINS be separated by gel electrophoresis?

A

By mass and charge

54
Q

What are DNA probes used for?

A

Identify the presence of a certain allele

55
Q

DNA probe definition

A

Short, single-stranded piece of DNA with a complementary base sequence to the allele

56
Q

How does a DNA probe identify a certain allele?

A

Binds the allele and have a radioactive/fluorescent label for detection

57
Q

Name the 6 stages of genetic screening

A
  1. Extraction
  2. Digestion
  3. Separation
  4. Southern blotting
  5. Hybridisation
  6. Autoradiography
58
Q

What happens in the extraction stage of genetic screening?

A
  1. DNA extracted from cell using homogenisation and centrifugation
  2. (PCR used to amplify DNA)
59
Q

What happens in the digestion stage of genetic screening?

A

DNA cut with restriction enzymes at recognition sites

60
Q

What happens in the separation stage of genetic screening?

A

DNA fragments separated by length and charge using electrophoresis

61
Q

What happens in the southern blotting stage of genetic screening?

A

DNA transferred from gel to a nylon membrane and made single-stranded

62
Q

What happens in the hybridisation stage of genetic screening?

A
  1. Complementary DNA probes added which bind to allele
  2. Membrane washed to remove any unbound probes and prevent false positives
63
Q

What happens in the autoradiography stage of genetic screening?

A

Fluoresence/radioactivity identified using photographic/X-ray film

64
Q

VNTR definition

A

Repeating DNA base sequences found BETWEEN genes

65
Q

How is genetic fingerprinting different to genetic screening?

A

Uses restriction enzymes and DNA probes complementary to VNTRs

66
Q

How are genetic fingerprints used in paternity tests?

A

All the bands in a child’s genetic fingerprint which don’t match the mother’s must match the father’s

67
Q

How are genetic fingerprints used in forensic science?

A

Genetic fingerprint of DNA found at a crime scene compared with the genetic fingerprint of a suspect

68
Q

How are genetic fingerprints used in animal and plant breeding?

A

Want to breed unrelated individuals to increase genetic diversity so check that there is variation between bands on genetic fingerprints

69
Q

How are genetic fingerprints used in medical diagnosis?

A

Genetic fingerprints of known types of tumour compared to genetic fingerprints of a patient’s tumour to diagnose it

Biology Y13 (13 decks)

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