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Biology A-level > Recombinant DNA technology > Flashcards

Recombinant DNA technology Flashcards

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1
Q

What is the function of restriction endonucleases?

A

To cut out specific sequences of DNA

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2
Q

What is the function of DNA ligase?

A

Joins together DNA sequences from different organisms

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3
Q

What is the importance of sticky ends?

A

If DNA is cut with the same restriction endonuclease it allows DNA from different organisms to be joined together via complementary base pairing.

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4
Q

What are the three ways of isolating a gene?

A

Reverse transcriptase
Restriction enzymes
Gene machine

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5
Q

What is PCR?

A

Polymerase chain reaction

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6
Q

What are the three stages of PCR?

A
  • Reaction mixture is set up with DNA fragment, primers, taq DNA polymerase and free nucleotides
  • DNA mixture is heated to break the hydrogen bonds. It is then cooled so the primers can anneal.
  • Reaction mixture is heated so DNA polymerase can work. DNA polymerase lines up complementary free nucleotides. Two new copies of DNA fragments are made.
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7
Q

Why is Taq polymerase useful?

A

It is thermostable so it does not need replacing every cycle

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8
Q

Compare PCR and semi-conservative replication

A

PCR only replicates short regions of DNA but SCR can replicate the whole genes.
PCR uses 95’C to split the strands and break hydrogen bonds but SCR uses semi conservative replication.
PCR uses primers, SCR does not.

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9
Q

What is proteome?

A

Full set of proteins produced by a certain genome

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10
Q

What do you need to do to DNA before you can sequence it?

A

Break it into fragments

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11
Q

How do you ensure the bacteria takes up the recombinant plasmid?

A
  • Mix recombinant plasmid and bacteria
  • Add calcium ions and heat making the bacteria more permeable
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12
Q

How can you identify which bacteria have taken up the plasmid?

A

Use a marker gene
This could be antibiotic resistance, fluorescent or an enzyme

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13
Q

What is a DNA probe?

A

Short, single stranded section of DNA with a specific sequence complementary to the base of interest.

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14
Q

How can a DNA probe be labelled?

A

Radioactively or fluorescently

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15
Q

What is genetic screening?

A

Analysing an individuals DNA for the presence of a particular gene

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16
Q

What are the five steps in DNA fingerprinting?

A
  • Extraction
  • Digestion
  • Separation
  • Hybridisation
  • Development
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17
Q

By what two principles does gel electrophoresis work?

A
  • Size (short moves faster)
  • DNA is negatively charged so travels towards the positive electrode
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18
Q

Why do we not create probes/primers that bind to only a triplet?

A

Same triplet occurs many times in genome, creating false positives

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19
Q

What do you need to do to a sample of DNA before you can sequence it?

A

Break it into fragements

20
Q

What did the human genome project do?

A

Mapped the whole human genome

21
Q

Why is it easier to determine the proteome of simple organisms?

A

There is very little non-coding DNA

22
Q

Who developed the first DNA sequencing technique?

A

Frederick Sanger

23
Q

What are some advantages of sequencing the genome and proteome of organisms?

A
  • identifying mutations in diseases
  • Identification of antigens during vaccine production
  • identification of new variants of a pathogen.
  • Finding out more about the evolutionary history of an organism
24
Q

What are organisms that contained transferred DNA known as?

A

Transgenic organisms

25
Q

What is the role of reverse transcriptase?

A

Produces DNA from RNA

26
Q

What is the role of protecting groups?

A

Makes sure the nucleotides are joined at the right points to prevent branching

27
Q

What does the term in vivo cloning mean?

A

Gene copies made inside a living organism

28
Q

What is a vector?

A

Something used to transfer DNA into a living cell

29
Q

What is the reaction mixture in step 1 of PCR?

A

DNA fragments, free nucleotides, primers and taq polymerase

30
Q

What is the role of a promoter region?

A

Determines whether a cell produces a protein or not

31
Q

What are the benefits of transformed organisms in agriculture?

A

Gives higher yields and crops have resistance to pests and droughts

32
Q

What are the benefits of transformed organisms in industry?

A

Reduces cost and can make food suitable for vegatarians

33
Q

What are the benefits of transformed organisms in medicine?

A

Drugs can be produced quicker and cheaper

34
Q

What are the disadvantages of transformed organisms in agriculture?

A

Monoculture
Super weeds
Organic farms can be contaminated

35
Q

What are the disadvantages of transformed organisms in medicine?

A

Use of life saving treatments could be limited and there are ownership issues once genetic material has been removed from the donors body

36
Q

What are the disadvantages of transformed organisms in industry?

A

Insufficient labelling and larger companies could take over

37
Q

What do both types of gene therapy involve?

A

Inserting a DNA fragment into a persons DNA

38
Q

Give three examples of vectors that can be used to insert a DNA fragment into a patients DNA

A
  • Altered virus
  • Plasmids
  • Liposomes
39
Q

What is somatic gene therapy?

A

Altering the alleles in a body cell

40
Q

Name a disease that can be treated using somatic gene therapy?

A

Cystic fibrosis

41
Q

What is germ line gene therapy?

A

Involves altering the alleles in sex cells

42
Q

What is personalised medicine?

A

Using genetic screening to determine which drug works best for a patient

43
Q

What is genetic counselling?

A

Advising patients and their relatives about the risk of genetic disorders. Can be advise on having a genetic screen, the results and how treatments work if the results are positive.

44
Q

State five uses of genetic fingerprinting?

A
  • Determining genetic relationships
  • Determining genetic variability
  • Forensic science
  • Medical diagnosis
  • Animal/plant breding
45
Q

What is a VNTR?

A

variable number tandem repeat.
Section of DNA that does not code for a protein. The length of these repeats at different locations varies from person to person.

46
Q

What is the role of fluorescent tag on DNA primers in genetic fingerprinting?

A

Used in the development stage when the gel has finished running. Gel is exposed to UV light and the banding pattern can be seen due to the fluorescent tags. This banding pattern can be compared to a selection of DNA fragments with known lengths.

Biology A-level (22 decks)

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