8 Answers to end-of-chapter questions

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Structured questions
10 a

Cutting the gene / length of DNA for insulin from genome

From mRNA carrying information for making insulin
From the amino acid sequence for insulin

Restriction / restriction endonucleases

[1]

ii

BamH1

[1]

Any 2 points [2]

iii
sticky ends
tetracycline-resistance gene

Good drawing showing correct cut [1]

Labels [1]
iv

Sticky ends produced / the terminal portion has a stretch of

unpaired nucleotides
the strands are not of the same length

[1]
[1]

[1]

DNA ligase
Is used to link together the deoxyribosephosphate backbones
of the DNA molecule producing a closed circle of the doublestranded DNA

Biology for CAPE

Original material Cambridge University Press 2011

[1]

vi

Adding of calcium ions to make the membrane more

permeable / by heat shock / adding calcium ions at 0 C and
raising it to 40 C suddenly
Or by electrophoresis

Any point [2]

vii Cloning of the treated bacteria

c

11 a

[1]

Screening of the bacteria to determine which have the

recombinant DNA
Multiplication / cloning of selected colony with insulin gene
Expression of gene / production of insulin
Adding a gene that causes fluorescence under ultraviolet light
Adding a gene that produces a harmless product that can be
stained

Any 2 points [2]

Any point [1]

plasmid rejoined,
tetracycline-resistance
gene rejoined

insulin gene joins

with itself to form
a closed circle

insulin gene
added to
plasmid,
forms closed
circle

For each
outcome drawing [1]
Labels /
explanations [1]
[max 6]

ii

Antibiotic resistance

[1]

iii

Exposed to ampicillin
The 3 black and 3 grey colonies survive exposure to
ampicillin
The bacteria would have taken up the plasmid with the
ampicillin-resistance gene
These bacteria will include recombinant and non-recombinant
plasmids
The circular foreign DNA which were taken up by some
bacteria (white colonies) did not have the ampicillinresistance gene and did not survive
1 point 1 mark [max 3]
Exposed to tetracycline
Any colonies that are resistant to tetracycline would not have
the foreign DNA
These colonies would survive / the grey colonies
If the foreign DNA is added to the plasmid, the tetracycline
gene would be disrupted
These colonies would die / the black colonies

Biology for CAPE

1 point 1 mark [max 4]

Original material Cambridge University Press 2011

Essay questions
12 a

13 a

Made in bacteria
Used to cut up viral DNA
To protect the bacteria from invading viruses / phages
Cut specific base sequences
Used in genetic engineering

5 points [3]
34 points [2]
12 points [1]

Obtain the required gene (cDNA) using mRNA carrying information for
making insulin and reverse transcriptase
Make the single-stranded cDNA double stranded and with sticky
ends
Use restriction enzyme to cut the vector plasmid / pBR322
Vector has 2 antibiotic-resistance markers
The tetracycline-resistance gene would be cut by restriction enzyme
Mix the cut vector plasmid with the insulin DNA fragments and
DNA ligase
Mix the plasmids / vector with the host cells / bacterial cells
Incubate in a calcium chloride solution to make bacterial cells more
permeable to the plasmids
Allow the bacteria to clone themselves
Screen the colonies to determine which ones have the insulin gene
Using replica plating, subject the colonies to ampicillin
1415 points [7]
The colonies that die do not have a vector / plasmid / antibiotic1213 points [6]
resistance markers
1011 points [5]
Subject the rest of the colonies to tetracycline
89 points [4]
The colonies that die contain the insulin gene since tetracycline gene
67 points [3]
is inactivated
45 points [2]
These colonies are cloned and the insulin gene expressed
23 points [1]
Advantage: simple procedure
Disadvantage: could increase the risk of other, potentially
pathogenic, bacteria taking up the genes and becoming resistant to
antibiotics
Cheaper
More efficient than collecting from animals
Identical to human insulin so better response / little immune reaction
Ethical / religious reasons for not collecting from animals

[2]

Any point [1]

[max 3]

DNA made by joining lengths of DNA from two different species

Organism which possesses DNA / genes from another species and
which is incorporated in their genome and actually expressed. They are
created to produce a new phenotype

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Benefits
The social impact has to do with its potential and actual impact on
human society and individuals
Higher crop yields so that people have more food
Enhance the nutritional content of crops (e.g. Golden Rice) so
that people are better fed / fewer deficiency diseases
Permit better targeted clean-up of wastes and pollutants

Biology for CAPE

Original material Cambridge University Press 2011

Lead to production of more effective and cheaper medicines

and treatments (e.g. insulin)

Potential hazards
Produce super-weeds or otherwise interfere with ecosystems in
unexpected ways, reducing crop yields so that people have less
food
Increase costs of seed
Some seeds cannot be used for sowing the next year since they
are modified to prevent germination thereby reducing crop
production / increasing debt burdens
Cause antibiotics to become less useful
Any well-discussed point [1]
Any valid point
[max 6]
c

14 a

Ethical impact is to do with issues of whether it is right or wrong, good

or bad to conduct research and develop genetically modified organisms
(GMOs)
[1]
It is good to conduct such research to develop GMOs that might
improve nutrition, the environment and health
It is wrong to continue such research when the potential impact of
the technology is unknown and many aspects of it remain to be
understood
It is wrong to use the results of such research when it involves
release of GMOs into the environment as once it is released it
cannot be taken back / there are unknown risks
Any valid point
Any 4 well described points [4]
The treatment of a disease by manipulating the genes in a persons
cells
Treatment of a genetic disorder by altering a patients genotype /
genome
Dominant allele can mask defective recessive allele when added to
the genome
Recessive allele does not have to be removed / inactivated / switched
off
Dominant disease allele would have to be inactivated which is very
difficult to do

Any 2 points [2]

Person suffering from CF has faulty CFTR protein

Chloride ion channels are defective
Results in thick mucus in respiratory passages
Faulty CFTR alleles are recessive single gene disorder
Somatic gene therapy employed
Normal allele is inserted into liposomes / vector
The vector / liposomes are introduced into patient by nasal sprays
The liposomes move through the plasma membrane carrying the
dominant allele
The allele must be incorporated into the affected cells / hosts
DNA
The dominant normal allele should be transcribed and then
translated
A normal protein would be produced
Symptoms should be relieved
Since it is somatic / cells die, treatment has to be repeated

1213 points [6]

1011 points [5]
89 points [4]
67 points [3]
45 points [2]
23 points [1]

Biology for CAPE

Any point [2]

Original material Cambridge University Press 2011

Benefits
Cures the patient / no longer suffers from the disease
Relieves some symptoms
Can now lead a normal life

Any 2 benefits [2]

Hazards
Allele may be inserted into another gene, altering the product /
disrupting its function
Allele may be inserted into tissue other than target tissue with
unknown consequences
Allele may be inserted into germ line and passed via gametes
Virus which carries allele may damage the tissue
Allele may be inserted into a promoter of a gene and may
interfere with gene switching
Allele may be inserted in a position where it affects the genes that
normally control the cell cycle
Any valid point

Any 3 hazards [3]

Biology for CAPE

Original material Cambridge University Press 2011