Question 1
Question
A radioactive probe can take what forms?
Answer
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ssDNA
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RNA
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Denatured dsDNA
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Oligonucleotides
Question 2
Answer
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Radioactive
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Fluorescent
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Use Ig's
Question 3
Question
Name the two ways probes can be designed
Question 4
Question
What is codon bias?
Answer
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Where cells prefer one codon of an aa to another
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Where cells will reject certain codons due to what they contain
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Where some cells can only make tRNA's for certain codons
Question 5
Question
80% of Phe codons are TTC, 20% are TTT
Question 6
Question
If the protein is an enzyme it can be found by adding the substrate
Question 7
Question
State the possible forms of Ig's for protein separation?
Answer
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Immobalised on a column
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Removable from solution
Question 8
Question
Probes needn't be 100% accurate to work
Question 9
Question
For a Western Blot, how is the DNA stuck to the membrane?
Question 10
Question
Once the DNA has been bound, what happens next?
Question 11
Question
In an expression vector, where is the new gene added?
Question 12
Question
Name the parts on the vector
Answer
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Origin
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Antibiotic Resistance
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Promoter
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Terminator
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Multiple Cloning Site
Question 13
Question
More compliated vectors have proteins tagged for purification
Question 14
Question
What is involved in His Chain Purification?
Answer
-
The addition of a His chain to the N terminus
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The removal of a His chain from the protein
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The use of a His chain to cut unwatned proteins
Question 15
Question
His Purification uses Histadine's affinity for what metal?
Question 16
Question
His-tagged proteins will bind to Nickel in a column. Others that don't bind will be washed off.
Question 17
Question
The PET system involves using a promoter for the protein that you can control the expression of
Question 18
Question
What promoter is used in the PET system to control protein expression?
Question 19
Question
In the PET system, what promoter is used to control the T_ expression?
Answer
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Lac promoter
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Slo Promoter
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Pap promoter
Question 20
Question
What is required for a good shuttle vector?
Question 21
Question
Shuttle vectors are usually plasmids with additional sequences
Question 22
Question
Shuttle vectors for yeast/fungi need
Question 23
Question
A baclovirus would be used as a vector into what?
Question 24
Question
Secretion of certain proteins can be caused by what?
Answer
-
Disulphide bonds
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Acetylation
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Methylation
Question 25
Question
Why are there two origins in the Yeast shuttle vector?
Question 26
Question
What is the method of selection in yeast?
Answer
-
URA3- allows growth in the absence of Uracil
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TRA3- allows growth in the absence of thymidine
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GRA3- allows growth in the absence of guanine
Question 27
Question
Like bacteria, yeast cannot take up linear DNA
Question 28
Question
What does a YAC contain that a plasmid does not?
Answer
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Non-useful genes
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Yeast centromere
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Yeast telomeres
Question 29
Question
Yeast can allow DNA to be directly inserted into its genome
Question 30
Question
How can yeast accept DNA into its genome?
Question 31
Question
In order to directly insert a gene, you must flank the gene with identical stretches of the part of the genome you want to insert it in
Question 32
Question
Plants would use an aglobacterium T1 plasmid
Question 33
Question
How do you replicate transformed plant cells?
Answer
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Grown them as plantlets
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Clone them
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Asexually reproduce them
Question 34
Question
Mammal shuttle vectors are the same as yeast vectors, but the yeast origin is replaced with what?
Answer
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An SV40 promoter
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A UG35 promoter
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A FT12 promoter
Question 35
Question
Retroviruses can also be used to insert transgenes into mammilian genomes by packing the transgene in a retroviral capsule