PRACTICAL QUESTIONS
1. Studies show that in plant leaves a pathway is not active because
one single enzyme in the pathway is not functional (while all
others are). The pathway can be restored by expressing a gene
encoding
a. a functional ortholog of this enzyme
b. this enzyme with a strong promoter
c. a transcription factor enabling expression of the pathway that this
enzyme is involved in
2. Anthocyanin expression is highly active in strawberry fruits, but
not in strawberry leaves. You can enable anthocyanin expression
in strawberry leaves by
a. Expressing all required enzymes in the strawberry leaves
b. Expressing the transcription factors that switch on all the genes of the
pathway in strawberry leaves
c. Expressing the first enzyme in the pathway
3. Can expression of the strawberry transcription factor(s) that
activate the anthocyanin pathway trigger this pathway in other
plants?
a. Yes in all plants as long as the transcription factors are expressed
constitutively
b. No you need to express all functional genes of the pathway
c. Yes but only in plants that carry the functional genes of the pathway
that can be activated by the strawberry transcription factor
d. No because the strawberry transcription factors will most likely not be
compatible with the anthocyanin genes of other plants
4. The FASTA sequence you just copied from NCBI was (1). Typical
of this type of sequence is that there are no (2) present.
1. a cDNA sequence of our GOI derived from the mRNA sequence,
a DNA sequence of our GOI derived from the plant genome
an mRNA sequence of our GOI derived from the plant's expressed
genes
2. No UTRS, introns, no exons, no introns
5. In the Gateway cloning system we insert AtMYB75 into a plasmid
using:
a. Recombination using the enzyme Clonase
b. Using restriction enzyme digestion and ligation
c. TOPO cloning
d. Blunt-end ligation
6. Which of the following schematic drawings best represents a PCR product
if the blue lines indicate an individual copy of the gene with the expected
sequence and the colored dots indicate a nucleotide that varies from the
expected sequence.
, 7. How should you adapt your PCR programme if the AT content of your
primers would be higher?
a. The denaturation temperature of the PCR programme should be lower
b. The denaturation temperature of the PCR programme should be higher
c. The denaturation time of the PCR programme should be shorter
d. The denaturation time of the PCR programme should be longer
e. The annealing temperature of the PCR programme should be lower
f. The annealing temperature of the PCR programme should be higher
g. The annealing time of the PCR programme should be shorter
h. The annealing time of the PCR programme should be longer
i. The elongation temperature of the PCR programme should be lower
j. The elongation temperature of the PCR programme should be higher
k. The elongation time of the PCR programme should be shorter
l. The elongation time of the PCR programme should be longer
8. How should you adapt your PCR programme if your gene would be twice
the size it is now?
a. The denaturation temperature of the PCR programme should be lower
b. The denaturation temperature of the PCR programme should be higher
c. The denaturation time of the PCR programme should be shorter
d. The denaturation time of the PCR programme should be longer
e. The annealing temperature of the PCR programme should be lower
f. The annealing temperature of the PCR programme should be higher
g. The annealing time of the PCR programme should be shorter
h. The annealing time of the PCR programme should be longer
i. The elongation temperature of the PCR programme should be lower
j. The elongation temperature of the PCR programme should be higher
k. The elongation time of the PCR programme should be shorter
l. The elongation time of the PCR programme should be longer
9. How is the DNA run on our agarose gel visualised? By incorporating (1) in
the agarose gel. This compound emits orange light when exposed to (2)
1. Bromethane/GelRed/EZ-vision/RNA
2. UV-light/bromo-phemol-orange
10. When you load DNA on gel, you use sample buffer that includes
glycerol and bromophenol blue. Why do you need these chemicals?
Glycerol:
a. gives your sample a higher density so that the sample quickly sinks and
remains in the slot after pipetting
b. binds to the DNA and gives it charge, which allows migration when a
current is applied.
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