Gene technology questions

Explain the use for fluorescent or easily stained substances markers in

gene technology (Ms answer) 8 marks gained in this answer
- Gene markers are inserted downstream the promoter region
beside the gene of interest, so when the gene of interest is
transcribed the gene marker is transcribed along with it
- Where gene markers such as GFP OR GUC, have ability of
fluorescing a visible colour such as green, when they are
exposed to ultraviolet rays
- This is easily identifiable, facilitating identification of the
genetically modified organism
- Where they have no harmful effect on the host cell

Explain why plasmids re frequently used in gene technology

1. They have a small piece of circular DNA, so they do not
contribute much to the DNA of host cell
2. They have ability to replicate independently from the host
cell
3. Have large number of recognition sites on the DNA for
restriction enzyme
4. Where DNA can be easily cut and added using DNA ligase
5. It can be easily extracted from bacteria and easily taken in by
bacteria
6. Have ability to act as vectors transferring substances from
one organism to another
7. They possess gene markers, where they have gene for
antibiotic resistance
8. They carry a promoter to allow expression of desired gene

How bioinformatics help identify the gene as suitable form to insert in

human
1. Bioinformatics is a stores and process data on DNA and proteins
2. Where it can be compared with the human DNA
 3. To make sure they are similar

Differences between gene expression in prokaryotes and eukaryotes

using eukaryotic and prokaryotic promoter
- The sequence of bases in eukaryotic promoters is different from
the sequence of bases in prokaryotic promoters
- As well as the RNA polymerase enzymes are different in
eukaryotic and prokaryotes
- So the RNA polymerase will not be able to recognize promoter
from different cell type
- Therefore transcription will not be initiated

Describe how the gene coding for human insulin can be obtained and
inserted into a plasmid vector 11 marks were gained in this answer
1. mRNA from B cells in the islets of Langerhans is extracted
2. where DNA primers are added to anneal to the mRNA as
reverse transcriptase can only work on double stranded
3. where reverse transcriptase enzyme uses the mRNA as a template
strand to form a cDNA single stranded
4. so the DNA polymerase is then used to create a complementary DNA
strand forming a double stranded DNA with sticky ends
5. where the plasmids are obtained using restriction endonuclease
enzyme
6. forming complementary sticky ends to the cDNA
7. the cDNA and the plasmid are mixed together
8. where the complementary sticky ends anneal to one another
forming hydrogen bonds
9. DNA ligase is then used to link the cDNA with the plasmid forming
phosphodiester bonds within the sugar phosphate back bone

Advantages of using theoretical models in research rather than

testing possible drugs in the laboratory
 1. It is cheaper, as no drugs or materials are purchased or
produced
2. It is less time consuming as computer software modelling can
easily be stimulated and modified to suit experimental
changes
3. No need to test drugs on humans or animals
4. No need for purchase of laboratories equipment’s or
resources

Suggest how a verity of flower that has gene for survival in dark
conditions could be produced from the existing flowers WITHOUT
USING GENE TEGNOLOGY
1. By selective breeding
2. Crossing the flower with a flower that is well adapted in dry
conditions
3. And then choose the offspring that has gained the genes for
surving in dark conditions
4. Repeat this for several generations

REMEMBER THAT THE RESTRICTION ENZYME HAS A

SPECIFIC RECOGNITION SITE, ya3ni it has a specific sequence of
bases at which it will cut
- What happens if a mutation (change is base sequence) occurs at
on of the sites which it recognizes
 The restriction enzyme will no longer be able to
recognize it
 Thus that part is not cut
 So this will lead to DNA fragment cut of different
size
Explain why the production of memory cells prevents gene
therapy from working in long-term chronic conditions such as
CF, as the insertion of the virus (vector) causes an increase in the
memory cells production ?

1. The repeat of treatment, causes a more rapid immune response

2. So the target cells which express the new/healthy protein
become short lived as they will be attacked by the immune
system
3. Thus this decreases the gene expression on re administration
Describe the problems associated with an immune response using
other types of vectors
- Naked DNA
o Has to be injected into the target cell
o It could be rapidly broken down
- Viruses
o Small so only small amount of DNA can be carried
o Low probability of integration into host genome
o Could cause mutations in the host DNA ‘
- Liposomes
o Have low ability to insert DNA into host cell