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Mutagenesis

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Year 2 Test sobre Mutagenesis, creado por gina_evans0312 el 29/12/2013.
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Resumen del Recurso

Pregunta 1

Pregunta
What is site directed mutation?
Respuesta
  • Where mutagenesis is directed towards a specific site
  • Where certain sites are more prone to mutagenesis than others
  • Where certain sites cannot be mutated and must be worked around

Pregunta 2

Pregunta
What form must the DNA be in for Site Directed Mutagenesis?
Respuesta
  • ssRNA
  • ssDNA
  • dsDNA

Pregunta 3

Pregunta
The primer for site directed mutagenesis is designed to have one or more mismatches
Respuesta
  • True
  • False

Pregunta 4

Pregunta
Once the mismatch has been achieved, how is it used?
Image:
Site_Directed (image/png)
Respuesta
  • During repair, the old base will be replaced with a new match
  • During replication, half the daughter cells will carry the mutation

Pregunta 5

Pregunta
What is specific about the DNA used in the Kunkel Method?
Respuesta
  • It's dsDNA
  • It's ssDNA
  • From the Slo + form of E.Coli
  • From the dut ung. form of E.Coli

Pregunta 6

Pregunta
What is Incorporated into the original DNA in order to make it susceptible to degrading
Respuesta
  • Uracil
  • Methyl-cytosine
  • Thymidine-3-phosphate

Pregunta 7

Pregunta
Like with site directed, the primer annealed to the DNA is designed to create mismatches
Respuesta
  • True
  • False

Pregunta 8

Pregunta
In both site directed and the Kunkel method, how is the primer extended?
Respuesta
  • DNA polymerase
  • Taq Polymerase
  • RNA polymerase

Pregunta 9

Pregunta
Once the DNA has been inserted, what happens to the original DNA strand?
Respuesta
  • The Uracil cause it to be partially degraded
  • The Methyl-cytosine cause it to be partially degraded
  • The Thymidine-3-Phosphate cause it to be partially degraded
  • Being removed alone with the bp next to them

Pregunta 10

Pregunta
The partially degraded strand is them re-synthesised by DNA repair proteins, finalising the mutation
Respuesta
  • True
  • False

Pregunta 11

Pregunta
Which method has a way of determining which plasmid has the mutation?
Respuesta
  • Site- Directed
  • Altered Site
  • Kunkel Method

Pregunta 12

Pregunta
What is particular about the plasmid used in the Altered Sites method?
Respuesta
  • It contains a non-functional Amp resistance gene
  • It contains a gene for DNA repair proteins
  • It's dsRNA

Pregunta 13

Pregunta
What oligonucleotide primers are used during Altered Site mutagenisis?
Respuesta
  • Mutant primer
  • Ampicillin 'Repair' primer
  • Ampicillin Non-Function primer

Pregunta 14

Pregunta
What is the result, after DNA replication has occurred?
Respuesta
  • The desired plasmid has the mutation, and tetracyline and ampicillin resistance
  • The desired plasmid has the mutation and tet resistance but no ampicillin resistance
  • The desired plasmid has the mutation and ampicillin resistance but no tetracyline resistance

Pregunta 15

Pregunta
What causes the mutagenesis in the 'Quickchange' PCR method?
Respuesta
  • A two primers containing the mutation
  • Rapid replication of the Site Directed mutant
  • One primer contains a mutation, the other is used to create the mutated plasmid
  • The original DNA is then degraded, leaving only the mutated plasmid

Pregunta 16

Pregunta
Why is Dpn used to degrade the original DNA?
Respuesta
  • It degrades only methylated DNA
  • It degrades only acetylated DNA
  • It degrades only ibiquitinated DNA

Pregunta 17

Pregunta
Site directed mutagenesis is unique in that no structural knowledge of the protein is required
Respuesta
  • True
  • False

Pregunta 18

Pregunta
State the two methods by which non-directed mutation occurs in evolution
Respuesta
  • Gradual accumulation of beneficiary mutations
  • Gradual loss of unnecessary DNA
  • Recombination of genes to form hybrid protein

Pregunta 19

Pregunta
These random mutations can be mimicked in a lab
Respuesta
  • True
  • False

Pregunta 20

Pregunta
Random mutagenesis works best by introducing many mutations at the same time
Respuesta
  • True
  • False

Pregunta 21

Pregunta
What will happen when this DNA replicates?
Image:
Degenerate_Primer (image/png)
Respuesta
  • The amino acid marked by the N's will be randomised
  • The Leu will be deleted
  • The N's are a stop codon, halting the gene

Pregunta 22

Pregunta
Primers can be more specific and limit the choice of potential aa further
Respuesta
  • True
  • False

Pregunta 23

Pregunta
Name an Error prone DNA polymerase
Respuesta
  • Taq
  • T7
  • Pol

Pregunta 24

Pregunta
Name the two conditions necessary for Error Prone mutagensis
Respuesta
  • An error prone polymerase
  • Conditions designed to increase error
  • No DNA repair enzymes
  • Restriction enzymes

Pregunta 25

Pregunta
DNA shuffling is used to create fusion proteins from two unrelated genes
Respuesta
  • True
  • False

Pregunta 26

Pregunta
Why does gene shuffling work?
Respuesta
  • Ligation repairs DNA errors
  • Related genes have overlapping areas
  • Related genes can ligate together with less error

Pregunta 27

Pregunta
What protein is used to randomly cut up two related genes for DNA shuffling?
Respuesta
  • DNAase I
  • DNAase III
  • EcoRV
  • EcoRI

Pregunta 28

Pregunta
The randomly cut up sequences are then ligated back together
Respuesta
  • True
  • False

Pregunta 29

Pregunta
When creating hybrids, the traditional approach of screening colonies for hybrids is not efficient- so we use robots
Respuesta
  • True
  • False

Pregunta 30

Pregunta
Name the two main methods of analysing mutants quickly
Respuesta
  • Surface display of proteins
  • Elution
  • Chemical compartmentalisation

Pregunta 31

Pregunta
The advantage of Protein Surface Display in improved mutations is?
Respuesta
  • Improved mutations will bind more strongly to immobalised substrate
  • They react more strongly to dyes
  • They don't degrade as easily

Pregunta 32

Pregunta
Once the variants are bound, they are washed so strongly only powerfully binding variants remain bound
Respuesta
  • True
  • False

Pregunta 33

Pregunta
Once the most improved variation has been identified, what can happen to it?
Respuesta
  • It's washed off and sequenced
  • It's washed off and improved again

Pregunta 34

Pregunta
Chemical Compartmentalisation uses an oil/acetyl emulsion
Respuesta
  • True
  • False

Pregunta 35

Pregunta
Why are the droplets important?
Respuesta
  • They make reading the DNA easier
  • Each droplet contains an individual mutation
  • Each droplet contains mutants that compliment each other

Pregunta 36

Pregunta
The DNA cannot be transcribed or translated in the droplets, only sequenced
Respuesta
  • True
  • False

Pregunta 37

Pregunta
Once the protein has been made, how are they assayed?
Respuesta
  • Robots put them on multi-well dishes for assay
  • Fluorescent activated cell sorters are used

Pregunta 38

Pregunta
How is fluorescence used for separation in Fluorescent Activated Cell Sorters?
Respuesta
  • The more fluorescent product is produced, the better the enzyme
  • The stronger the enzyme-substrate binding, the better the enzyme
  • The more fluorescent substrate is used, the better the enzyme
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