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Biology 2 > 10. Manipulating genomes > Flashcards

10. Manipulating genomes Flashcards

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1
Q

What are the benefits of knowing the human Genome?

A

Comparisons between species, Evolutionary relationships, Trace human migration, Variation between individuals, Epigenetics

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2
Q

What is epigenetics?

A

Understanding the developments of certain diseases.

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3
Q

What are the steps of sequencing DNA?

A
  1. Digestion of DNA using restriction enzymes.
  2. Polymerase chain Reaction
  3. Gel electrophoresis
  4. DNA sequencing
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4
Q

What are restriction enzymes?

A

They recognise a specific site along the length of DNA.

They then cut the strand at its recognition site.

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5
Q

What is palindromic DNA?

A

Restriction enzymes can read DNA in both directions.

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6
Q

What are the two ways that restriction enzymes cut DNA?

A

The restriction enzyme can cut the sequence in two different areas on each strand resulting in sticky ends.
Or the restriction enzyme can cut straight through the strand resulting in blunt ends.

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7
Q

What is gel electrophoresis?

A

Used to separate the fragments of DNA.

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8
Q

What are the stages of gel electrophoresis?

A
  1. A gel containing a buffer solution is made.
  2. Blue loading dye is added to the digested DNA.
  3. DNA is loaded into wells at the negative electrode.
  4. Connected to 18V and allowed to run for 6-8 hours.
  5. DNA is attracted to the positive electrode so moves through the gel.
  6. Larger fragments are slower to move while the smaller fragments move quicker.
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9
Q

How are the results of gel electrophoresis read?

A

The DNA produces a banding pattern which can be seen under a UV light due to the blue dye.
The patterns produced from different samples of DNA can then be compared.

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10
Q

What is the role of a DNA probe?

A

A short sequence that finds a corresponding sequence of DNA to the gene being looked for.

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11
Q

What is a DNA microarray?

A

Specific probes are placed on a fixed surface and DNA is applied.
The DNA is labelled with fluorescent markers which shows up under a scan if it binds to a particular probe. This can reveal the presence of mutated alleles.

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12
Q

What is DNA sequencing?

A

Determining the exact order of nucleotides in a segment of DNA.

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13
Q

What is PCR?

A

Artificial replication of short sequences of DNA.

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14
Q

What are the steps of PCR?

A
  1. A primer molecule, DNA polymerase and DNA base pairs are added to a length of DNA.
  2. Cycle of heating and cooling separate DNA strands, bind primers and replicate DNA strands.
  3. This can be repeated many times to create more copies of DNA.
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15
Q

What is the role of primers?

A

Allow DNA polymerase to attach to the length of DNA.

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16
Q

What are the uses of PCR?

A

Detecting mutations, Identifying viral infections, Monitoring the spread of infectious disease, Forensic science, Research into the genome of ancient human remains.

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17
Q

How do you calculate exponential Growth?

A 
N= Nᵒ x 2ᴺ
Nᵒ = Initial number
ᴺ= number of divisions, or generations
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18
Q

How do you represent the answer as a log₁?

A

Place the value into the calculator.
Press log₁₀.
Write the value as 10ᴺ.

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19
Q

What are the steps of the sanger method?

A
  1. Starts with a single-stranded DNA sequence, broken into chunks of different lengths and tagged.
  2. The final base ensures that no more can be added.
  3. The DNA fragments are passes through a gel by electrophoresis.
  4. This arranges the fragments in order of length to generate the DNA sequence.
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20
Q

What is Pyrosequencing?

A

Used to sequence DNA by using chemiluminescent enzymatic reactions.

21
Q

What are the steps of pyrosequencing?

A
  1. A length of DNA is mechanically cut into fragments.
  2. These lengths are then degraded into single-stranded DNA (template DNA) and are immobilised.
  3. A sequencing primer is added and the DNA is then incubated with certain enzymes.
  4. One of the four possible activated nucleotides is added at one time and any light generated is detected.
  5. This determines which base is added at each step to generate a sequence.
22
Q

What is an activated nucleotide?

A

A nucleotide with two extra phosphoryl groups.

23
Q

What is bioinformatics?

A

The storage of collected genome data on computers.

24
Q

What are tandem repeats?

A

Repetitive segments of DNA that do not code for proteins.

25
Q

What is DNA profiling?

A

The comparison of individuals profiles of DNA.

26
Q

What are the steps of DNA profiling?

A
  1. DNA is obtained from the individual
  2. The DNA is digested by restriction enzymes.
  3. Fragments are separated by gel electrophoresis.
  4. A banding pattern can be seen.
  5. The DNA to which the individual is being compared goes through the same process.
  6. The banding patterns are compared.
27
Q

What genetic information is used in DNA profiling?

A

Short tandem repeat (STR) sequences of DNA.

Highly variable short repeating lengths of DNA.

28
Q

What are the applications of DNA profiling?

A

Forensic science, Maternity and paternity disputes, Analysis of disease

29
Q

What are the steps of heating and cooling during PCR?

A
  1. Denaturation, in which double-stranded DNA is heated to separate the strands.
  2. Annealing, allows DNA primers to bind to the target region of DNA.
  3. Extension, DNA polymerase uses the DNA nucleotides to amplify the DNA using each strand as a template.
30
Q

What is genetic engineering?

A

Combining DNA from different organisms, resulting in the expression of the new gene.
Makes recombinant organisms described as transgenic.

31
Q

What is pharming?

A

Transgenic animals and plants modified to produce useful pharmaceuticals.

32
Q

What are the steps of genetic engineering?

A

The required gene is obtained.
A copy of the gene is placed inside a vector.
The vector carries the gene into a recipient cell.
The recipient expresses the novel gene.

33
Q

How is the required gene obtained for genetic engineering?

A
  1. mRNA from a cell is isolated.
  2. The enzyme reverse transcriptase and a supply of free DNA nucleotides are added.
  3. A single strand DNA is synthesised using the mRNA as a template.
  4. The enzyme ribonuclease H breaks down the mRNA strand.
  5. DNA polymerase and DNA nucleotides are added, synthesising a complementary strand of DNA.
34
Q

How is a gene inserted into a vector?

A
  1. A restriction enzyme will cut the plasmid at specific recognition sites, exposing sticky ends.
  2. Free nucleotide bases are added, forming a recombinant plasmid.
  3. DNA ligase catalysis the annealing (binding) of the gene and plasmid.
35
Q

What is a vector?

A

An agent that carries piece of DNA from one cell to another.

36
Q

What are the various methods used to aid DNA in crossing the cell’s plasma membrane?

A

Heat shock treatment
Electroporation
Electrofusion

37
Q

What percentage of bacterial cells take up plasmid?

A

<1%

38
Q

What are the three possible outcomes when inserting a plasmid into a bacteria cell?

A

Bacteria do not have the plasmid in them.
Bacterial that have taken up the plasmid, but the plasmid resealed without the gene.
Bacteria that have taken up the plasmid with the gene in it.

39
Q

How do bacteria cells react to ampicillin when a plasmid is and isn’t present?

A

When the plasmid is present within the cell, it has a resistance to ampicillin.

40
Q

How do bacteria cells react to tetracycline when the gene has and hasn’t incorporated with the plasmid in the cell?

A

When the plasmid with the gene incorporated is present within the cell, its resistance to tetracycline is disrupted.

41
Q

What are the steps of identifying a recipient bacteria cell that expresses the novel gene?

A
  1. The bacteria are cultured on agar plates with ampicillin. Those that did not accept the plasmid die.
  2. The remaining bacteria is then cultured on agar plates with tetracycline. The bacteria that have incorporated the plasmid with the gene inside will die.
  3. The bacteria that grew on the ampicillin but died on the tetracycline will be selected and replicated.
42
Q

What is Xenotransplantation?

A

The transfer of organs or tissues into humans from other species.

43
Q

What is golden rice?

A

Rice that has been engineered to contain beta-carotene which is converted into Vitamin A when digested.

44
Q

What are the beneficial uses of Genetic engineering?

A

Making pest and drought resistant crops especially for the developing world.
Making nutritionally enhanced crops.
Using transgenic animals to express pharmaceutical products such as alpha-antitrypsin to treat emphysema.
Using microorganisms to make treatments such as insulin.

45
Q

What are the concerns surrounding genetic engineering?

A

Microorganisms could escape into the wild and transfer antibiotic resistance to other bacteria.
The gene for herbicide resistance to pass into weeds, producing ‘superweeds’.
Worry that inserted genes in food will somehow be expressed in us.
Concerns for the welfare of the GM sheep and goats.

46
Q

What is Gene therapy?

A

When the cause of the genetic disease is targeted, rather than just the symptoms.
A functional allele of a particular gene is inserted into cells, masking the effects of the defective or mutated allele.

47
Q

What is the outcome if gene therapy is successful?

A

If this allele is expressed, then the individual will produce a functioning protein and no longer have the symptoms associated with the genetic disorder.

48
Q

What are the different methods of inserting functioning genes to the patient’s body cells?

A

Liposomes, Viruses, Somatic cell gene therapy, Germ line therapy

Biology 2 (12 decks)

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