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Mutagenesis

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Year 2 Quiz on Mutagenesis, created by gina_evans0312 on 29/12/2013.
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Quiz by gina_evans0312, updated more than 1 year ago
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Created by gina_evans0312 over 10 years ago
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Resource summary

Question 1

Question
What is site directed mutation?
Answer
  • 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

Question 2

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

Question 3

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

Question 4

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

Question 5

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

Question 6

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

Question 7

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

Question 8

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

Question 9

Question
Once the DNA has been inserted, what happens to the original DNA strand?
Answer
  • 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

Question 10

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

Question 11

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

Question 12

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

Question 13

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

Question 14

Question
What is the result, after DNA replication has occurred?
Answer
  • 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

Question 15

Question
What causes the mutagenesis in the 'Quickchange' PCR method?
Answer
  • 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

Question 16

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

Question 17

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

Question 18

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

Question 19

Question
These random mutations can be mimicked in a lab
Answer
  • True
  • False

Question 20

Question
Random mutagenesis works best by introducing many mutations at the same time
Answer
  • True
  • False

Question 21

Question
What will happen when this DNA replicates?
Image:
Degenerate_Primer (image/png)
Answer
  • 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

Question 22

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

Question 23

Question
Name an Error prone DNA polymerase
Answer
  • Taq
  • T7
  • Pol

Question 24

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

Question 25

Question
DNA shuffling is used to create fusion proteins from two unrelated genes
Answer
  • True
  • False

Question 26

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

Question 27

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

Question 28

Question
The randomly cut up sequences are then ligated back together
Answer
  • True
  • False

Question 29

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

Question 30

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

Question 31

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

Question 32

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

Question 33

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

Question 34

Question
Chemical Compartmentalisation uses an oil/acetyl emulsion
Answer
  • True
  • False

Question 35

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

Question 36

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

Question 37

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

Question 38

Question
How is fluorescence used for separation in Fluorescent Activated Cell Sorters?
Answer
  • 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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