Name: ________________________

3.8.4 Genetic Technologies Class: ________________________

Date: ________________________

Time: 385 minutes

Marks: 311 marks

Comments:

Page 1 of 92
 (a) Describe how a gene can be isolated from human DNA.
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(2)

(b) Describe how an isolated gene can be replicated by the polymerase chain reaction (PCR).

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(c) (i) Describe how a harmless virus, genetically engineered to contain a CFTR gene, can
be used to insert the gene into a cystic fibrosis sufferer.

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(2)

Page 2 of 92
(ii) A virus used in gene therapy has RNA as its genetic material and has an enzyme
called reverse transcriptase. Inside a human cell, reverse transcriptase uses viral
RNA to make viral DNA.

Explain why the enzyme is called reverse transcriptase.

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(1)
(Total 9 marks)

Page 3 of 92
 (a) Agrobacterium is a bacterium used in genetic engineering of plants. The diagram shows
2 stages in the transfer of a gene into a plant.

Page 4 of 92
 (i) Name structure X in stage 1.

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(1)

(ii) In stage 2, explain why the bacteria are cultured before the plant tissue is added.

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(iii) In stage 4, explain why the growth medium contains antibiotic.

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(iv) Suggest why stages 5 and 6 are necessary for the commercial production of
genetically engineered plants.

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(2)

(b) (i) A toxin that kills insects can be sprayed directly onto the leaves of crop plants. A
gene has now been transferred into crop plants that makes their leaves produce this
toxin.

Explain one advantage to farmers of growing the genetically engineered crop plants,
rather than spraying leaves with the toxin.

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(1)

Page 5 of 92
(ii) Suggest one reason why some people are concerned that the toxin gene might get
transferred to wild plants that are related to the crop plants.

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(1)
(Total 8 marks)

Page 6 of 92
 Silkworms secrete silk fibres, which are harvested and used to manufacture silk fabric.
3
Scientists have produced genetically modified (GM) silkworms that contain a gene from a spider.

The GM silkworms secrete fibres made of spider web protein (spider silk), which is stronger than
normal silk fibre protein.

The method the scientists used is shown in the figure below.

(a) Suggest why the plasmids were injected into the eggs of silkworms, rather than into the
silkworms.

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Page 7 of 92
(b) Suggest why the scientists used a marker gene and why they used the EGFP gene.

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(2)

The scientists ensured the spider gene was expressed only in cells within the silk glands.

(c) What would the scientists have inserted into the plasmid along with the spider gene to
ensure that the spider gene was only expressed in the silk glands of the silkworms?

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(1)

(d) Suggest two reasons why it was important that the spider gene was expressed only in the
silk glands of the silkworms.

1 .....................................................................................................................

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2 .....................................................................................................................

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(2)
(Total 7 marks)

Page 8 of 92
 Research scientists can increase the nutritional value of potatoes by genetically engineering
4 potato plants. A gene which results in increased protein production has been removed from cells
of an amaranth plant and inserted into cells of a potato plant.

(a) Describe how a gene could be removed from cells of an amaranth plant and inserted into
cells of a potato plant.

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(6)

(b) Whole potato plants can be produced from genetically identical potato cells grown in a
tissue culture. Use your knowledge of genes to suggest how different cells, such as leaf
and root cells, can develop from genetically identical cells.

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(2)
(Total 8 marks)

Page 9 of 92
 (a) Plasmids can be modified by genetic engineering and inserted into bacteria. These
5 bacteria can then make useful substances normally made by another organism. Explain
how modified plasmids are made by genetic engineering and how the use of markers
enable bacteria containing these plasmids to be detected.

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(b) In gene therapy, genes are introduced into a person who has defective genes which do not
produce an important substance. Three experiments were done to compare techniques for
introducing an important substance into a person with defective genes.

1. The substance was injected directly.

2. Harmless viruses carrying genes coding for the substance were injected.
3. The genes were put into a protein capsule which was inserted into the tissues.

Page 10 of 92
The graph shows results of the experiments.

Takahiro Ochiya et al, Biomaterials for Gene Delivery: Studies on Metastasis,

(National Cancer Centre, Research Institute, Tokyo, Japan) 1999

(i) Describe the results of the three experiments.

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(3)

(ii) Using the information in the graph, suggest one advantage and one disadvantage of
the capsule method compared to the others.

Advantage ...........................................................................................

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Disadvantage ............................................................................….......

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(2)
(Total 11 marks)

Page 11 of 92
 (a) Explain the reason for each of the following in the polymerase chain reaction (PCR).
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(i) DNA is heated to 95 °C.

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(ii) DNA polymerase used is heat-stable.

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(iii) The reaction mixture is cooled to 40 °C.

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(1)

(b) The graph shows the number of DNA molecules made using PCR, starting with one
molecule.

(i) Explain the shape of the curve from cycles 1 to 16.

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Page 12 of 92
 (ii) Suggest one explanation for the levelling out of the curve from cycles 17 to 20.

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(Total 7 marks)

The polymerase chain reaction is a process which can be carried out in a laboratory to replicate
7 DNA. The diagram shows the main stages involved in the polymerase chain reaction.

(a) Explain why DNA is heated to 95 °C.

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(1)

(b) What is the role of

(i) a primer in this process;

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(ii) DNA polymerase?

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Page 13 of 92
(c) (i) How many DNA molecules will have been produced from one molecule of DNA after
6 complete cycles?

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(1)

(ii) Suggest one use of the polymerase chain reaction.

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(1)

(d) Give two ways in which the polymerase chain reaction differs from the process of
transcription.

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(2)
(Total 7 marks)

Page 14 of 92
 (a) Cystic fibrosis can be caused by any one of several mutant alleles of the cystic fibrosis
8 gene. The most common of these mutant alleles accounts for about 70% of cases of cystic
fibrosis. The use of gene probes can identify individuals carrying this allele. Gene probes
are single strands of DNA which are radioactively labelled. They have a base sequence
that is complementary to a mutant allele. The main stages in using a gene probe are shown
in the diagram.

Sample of DNA extracted from a

person’s tissue and heated to
separate the strands

↓
Radioactive gene probe added
to the DNA

↓
Excess probe washed away

↓
Sample tested for radioactivity

Using the information given, explain how the use of a gene probe could enable the
presence of a mutant allele of the cystic fibrosis gene to be detected.

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Page 15 of 92
 (b) Sheep have been genetically engineered to produce alpha-1-antitrypsin which is used to
treat cystic fibrosis. Use your knowledge of this process to explain one argument for and
one against using sheep in this way.

For

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Against

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(2)
(Total 6 marks)

Gene therapy is used to treat the genetic disorder, ADA deficiency. Affected individuals are
9 unable to produce the enzyme adenosine deaminase (ADA). Without this enzyme, T
lymphocytes, a type of white blood cell, cannot provide immunity to infection. The diagram shows
the processes involved in the treatment of ADA deficiency by gene therapy.

(a) What is meant by gene therapy?

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(1)

Page 16 of 92
(b) The ADA gene is inserted into a virus. Give two advantages of using a virus in gene
therapy.

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(2)

(c) Individuals who have been treated by this method of gene therapy do not pass on the ADA
gene to their children. Explain why.

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(1)

(d) T lymphocytes are produced in bone marrow. A bone marrow transplant from a genetically
matched donor can provide a permanent cure for ADA deficiency.

(i) Suggest why bone marrow for a transplant is obtained from a genetically matched
donor.

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(1)

(ii) Explain why treatment of ADA deficiency by gene therapy must be repeated at
regular intervals, whereas a single bone marrow transplant can provide a permanent
cure.

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(2)
(Total 7 marks)

Page 17 of 92
 The polymerase chain reaction (PCR) can be used to produce large quantities of DNA.
10 Describe how the PCR is carried out.

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(Total 6 marks)

A protein produced by a species of bacterium is toxic to caterpillars. The gene coding for this
11 protein was removed and transferred into a crop plant.

(a) (i) Describe how the gene could have been removed from the bacterial DNA.

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(2)

(ii) Many copies of the isolated gene were required. Name the process used in a
laboratory to produce many copies of DNA from a small amount.

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(1)

Page 18 of 92
 (b) The gene was injected into isolated cells from the crop plant. These cells were then cloned
and new plants grown from the cloned cells. Explain the advantage of inserting the gene
into isolated plant cells rather than directly into cells within a whole plant.

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(Total 6 marks)

(a) Plasmids are often used as vectors in genetic engineering.

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(i) What is the role of a vector?

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(1)

(ii) Describe the role of restriction endonucleases in the formation of plasmids that
contain donor DNA.

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(iii) Describe the role of DNA ligase in the production of plasmids containing donor DNA.

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Page 19 of 92
(b) There are many different restriction endonucleases. Each type cuts the DNA of a plasmid
at a specific base sequence called a restriction site. The diagram shows the position of four
restriction sites, J, K, L and M, for four different enzymes on a single plasmid. The
distances between these sites is measured in kilobases of DNA.

1 kb = 1 kilobase

The plasmid was cut using only two restriction endonucleases. The resulting fragments
were separated by gel electrophoresis. The positions of the fragments are shown in the
chart below.

(i) Which of the restriction sites were cut?

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(1)

Page 20 of 92
 (ii) Explain your answer.

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(1)
(Total 6 marks)

DNA probes may be used to identify the presence of specific genes associated with human
13 diseases. The flow chart summarises the way in which they are used.

Stage 1 DNA is cut into fragments

Stage 2 Electrophoresis separates the DNA fragments

Stage 3 Radioactive DNA probes are used to locate specific DNA fragments

(a) Name the enzyme used in Stage 1.

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(1)

(b) Explain how electrophoresis separates the fragments of DNA in Stage 2.

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(c) (i) What is a DNA probe?

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Page 21 of 92
 (ii) Explain why radioactive DNA probes are used to locate specific DNA fragments.

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(2)
(Total 7 marks)

Read the following passage.

14
Malaria is a disease so deadly that it has devastated armies and destroyed great
civilisations.
It has been estimated that in the course of history malaria has been responsible for the death
of one out of every two people who have ever lived. Even today, with all the advantages of
modern technology, it is still responsible for some three million deaths a year.

5 The first half of the twentieth century was a time of hope for malarial control. The drugs
chloroquine and proguanil had just been discovered and there seemed a real possibility of a
malaria-free world. Unfortunately, this honeymoon ended almost as soon as it had started,
with the emergence of drug-resistant parasite populations. Scientists now accept that
whatever
new drug they come up with, it is likely to have a very limited effective life. As a result, they
10 are increasingly looking at combinations of drugs.

The approach to malaria control which holds the best hope is the production of a vaccine.
One
of these is being developed by a researcher in South America. His vaccine is based on a
small
synthetic polypeptide called SPf66 which is dissolved in a saline solution and given as an
injection. A series of early trials on human volunteers produced confusing results. In one trial
15 the effectiveness of the vaccine was claimed to be 80% while, in others, the results were
statistically insignificant. Not only were the results inconclusive but the methods used were
challenged by other scientists. In particular, the controls were considered inappropriate.

Another, possibly more promising, approach has been the development of a DNA-based
vaccine. In theory, all that is required is to identify the DNA from the parasite which encodes
20 key antigens. Unfortunately, scientists have hit snags. Although they have succeeded in
sequencing the human genome, the genome of the malarial parasite has created major
difficulties. This is partly because of the very high proportion of the bases adenine and
thymine. In some places these two bases average 80%, and on chromosomes 2 and 3
nearly
100% of the bases present are adenine and thymine. Because of this, it has proved
impossible
25 to cut the relevant DNA with the commonly available restriction enzymes into pieces of a
suitable size for analysis.

Page 22 of 92
Use information from the passage and your own knowledge to answer the following questions.

(a) Explain how a resistant parasite population is likely to arise and limit the life of any new
anti-malarial drug (lines 8 - 9).

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(b) A person has a 1 in 500 probability of being infected by a chloroquine-resistant strain of

malarial parasite and a 1 in 500 probability of being infected by a proguanil-resistant strain.
Use a calculation from these figures to explain why scientists are “increasingly looking at
combinations of drugs” (lines 9 - 10).

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(c) (i) Explain why trials of the SPf66 vaccine needed a control.

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(1)

(ii) The controls for the SPf66 vaccine trials were considered inappropriate (line 17).

Suggest how the control groups in these trials should have been treated.

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Page 23 of 92
(d) In some of the DNA of a malarial parasite, the proportion of adenine and thymine bases
averages 80% (lines 22 - 23). In this DNA what percentage of the nucleotides would you
expect to contain

(i) phosphate; ..........................................................................................

(ii) guanine? .............................................................................................

(2)

(e) (i) Use your knowledge of enzymes to explain why restriction enzymes only cut DNA at
specific restriction sites.

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(3)

(ii) Restriction enzymes that can cut the DNA of chromosomes 2 and 3 produce pieces
that are too small for analysis. Explain why these restriction enzymes produce small
DNA fragments.

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(2)
(Total 15 marks)

Page 24 of 92
 ‘Take-all’ is a disease of wheat caused by a fungus. It can cause serious damage to the crop.
15
There is no gene for resistance to this fungus in wheat. There is, however, a gene for resistance
to this fungus present in oats.

The diagram shows how this gene might be transferred to wheat.

(a) (i) The wheat plant with the resistance gene contains recombinant DNA. What is
recombinant DNA?

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(1)

(ii) The plasmids act as vectors for the resistance gene. What is a vector?

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Page 25 of 92
 (iii) Suggest how cells with the resistance gene might be selected.

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(2)

(b) A laboratory has oat plants containing the resistance gene and a supply of plasmids.

Describe how bacteria may be produced which have the resistance gene in their plasmids.

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(Total 10 marks)

Page 26 of 92
 Read the following passage.
16
In 1991, nine skeletons were found in Russia. They were believed to be those of Tsar Nicholas II,
his family and staff who were killed in 1917 during the Russian revolution. Very small amounts of
DNA were isolated from these skeletons. This DNA was used in the polymerase chain reaction
(PCR). Genetic fingerprinting was then carried out on this DNA to identify the skeletons.

The chart shows some of the results obtained from the genetic fingerprinting of seven of the
skeletons, three children and four adults.

Use information from the passage and your own knowledge to answer the following questions.

(a) Explain why the polymerase chain reaction was used in this investigation.

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Page 27 of 92
(b) In the polymerase chain reaction, DNA is heated to 95 °C and nucleotides, enzymes and
DNA primers are added to the mixture.

(i) Explain why the DNA is heated to 95 °C.

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(1)

(ii) What are DNA primers?

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(1)

(iii) Why are DNA primers added during the polymerase chain reaction?

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(1)

(iv) What is the advantage of the enzyme used in the polymerase chain reaction being
thermostable?

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(2)

Page 28 of 92
(c) Describe how genetic fingerprinting is carried out.

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(6)

(d) All three children on the chart had the same parents. One of the parents was Adult 1.

Which of the other three adults on the chart was the other parent? Give the reason for your
answer.

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(2)
(Total 15 marks)

Page 29 of 92
 (a) (i) Some human DNA was cut into separate pieces using a restriction enzyme which
17 produced a staggered cut. A scientist wanted to insert these pieces of DNA into
plasmids and used the same restriction enzyme to cut the plasmids. Explain why the
pieces of human DNA would be able to join to the cut DNA of the plasmids.

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(2)

(ii) Which other enzyme must the scientist have added to the mixture to form
recombinant plasmids?

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(1)

(b) A plasmid may be used as a vector. Explain what is meant by a vector in this context.

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(2)

(c) Molecular biologists often use plasmids which contain antibiotic resistance genes.

Explain the reason for this.

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(2)
(Total 7 marks)

Page 30 of 92
 β-thalassaemia is a genetic condition in which abnormal haemoglobin is produced. In one form,
18 the recessive allele for β-thalassaemia, t, differs from the normal allele, T, by a single base-pair.
A radioactive DNA probe was used to investigate the genotypes of four members of one family.
The flowchart summarises the technique involved.

DNA samples extracted and cut into fragments using a

restriction enzyme

↓
Fragments separated from each other by electrophoresis

↓
One region of the resulting gel was blotted with two pieces
of filter paper. The first was soaked in a solution containing
a radioactive DNA probe for the normal allele.
The second was soaked in a solution containing a
radioactive DNA probe for the β-thalassaemia allele.

↓
Surplus probe washed off

The diagram below shows the appearance of the two pieces of filter paper which resulted from
the investigation.

(a) What is the probability that the next child that this couple have is a girl who has
β-thalassaemia? Explain your answer.

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Page 31 of 92
 (b) (i) The fragment of DNA containing the normal allele and the fragment with the
β-thalassaemia allele moved the same distance on the gel. Explain why.

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(2)

(ii) The allele for β-thalassaemia differs from the normal allele by only one base-pair.
Explain why the probe used to identify these alleles consists of a piece of DNA
twenty bases in length and not just one base.

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(2)
(Total 7 marks)

(a) (i) Some human DNA was cut into separate pieces using a restriction enzyme which
19 produced a staggered cut. A scientist wanted to insert these pieces of DNA into
plasmids and used the same restriction enzyme to cut the plasmids. Explain why the
pieces of human DNA would be able to join to the cut DNA of the plasmids.

.............................................................................................................

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(2)

(ii) Which other enzyme must the scientist have added to the mixture to form
recombinant plasmids?

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(1)

Page 32 of 92
(b) A plasmid may be used as a vector. Explain what is meant by a vector.

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(2)

(c) Molecular biologists often use plasmids which contain antibiotic resistance genes.

Explain the reason for this.

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(2)
(Total 7 marks)

Page 33 of 92
 Read the following passage.
20
Shark-fin soup is an expensive delicacy. To provide the basic ingredient, fishermen catch the
sharks, hack the fins off and throw the dead bodies back into the ocean. But sharks are slow
to mature and produce only a few offspring at a time, so they are vulnerable to overfishing.
Monitoring the shark-fin trade is difficult, as once a fin has been cut off, it can be extremely
5 difficult to work out precisely from which species it was taken.

The DNA from different species of sharks shows some differences in base sequence. This
has
enabled a new genetic fingerprinting technique to be developed. This technique would allow
conservationists and fisheries managers to assess which of the 400 shark species are most
threatened by the trade in shark fins.

10 An identification process has been developed using a range of “primers”. These are short
pieces of single-stranded DNA that are complementary to a particular sequence of DNA.
Each primer is specific to the DNA of one shark species.

The primers are added to DNA taken from a shark’s fin and the polymerase chain reaction is
carried out. Only two primers, one at each end of a certain piece of DNA, will bind. The
piece
15 of DNA between the primers is replicated by the polymerase chain reaction. The primers that
bind are specific to a particular species of shark and the length of the DNA fragment
replicated differs for each species. When this DNA is run in an electrophoresis gel it
produces
a single band, enabling the researchers to identify which species of shark is involved.

Use information from the passage and your own knowledge to answer the questions.

(a) (i) Explain why the DNA for each species of shark shows differences in base sequence
(line 6).

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(2)

(ii) Each primer is specific to the DNA of one shark species (line 12).

Explain why a particular primer will only bind to the DNA of one species.

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Page 34 of 92
 (iii) The length of the replicated DNA fragment is different for each species.

Explain why this is important in identifying the shark species involved.

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(3)

(b) In conventional DNA fingerprinting, a series of bands is produced on the electrophoresis

gel, resembling the rungs of a ladder. When the DNA in this new genetic fingerprinting
technique is run in an electrophoresis gel it produces just one of these ‘rungs’.

Explain the reason for the difference in the number of ‘rungs’ produced.

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Page 35 of 92
 (c) Describe the polymerase chain reaction.

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(6)
(Total 15 marks)

Scientists are working to produce a genetically modified bacterium to treat patients suffering from
21 a disease of the digestive system. They plan to collect mRNA from human cells. This will be used
to produce the DNA of the gene for the protein interleukin. They will then transfer this human
gene into the bacterium Lactococcus. The scientists intend patients to swallow the genetically
modified bacteria. These bacteria will release interleukin inside the digestive system to treat the
disease.

(a) (i) Name the type of enzyme which will be used to produce the DNA from the mRNA.

.............................................................................................................
(1)

(ii) It is easier to obtain the interleukin gene from mRNA rather than directly from the
DNA removed from human cells. Explain why.

.............................................................................................................

.............................................................................................................
(1)

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 (b) The scientists propose to put the gene directly into the DNA of Lactococcus.
Describe the role of the enzyme ligase in this process.

......................................................................................................................

......................................................................................................................
(1)
(Total 3 marks)

Read the following passage.

22
DNA tests were used to confirm the identity of deposed Iraqi leader Saddam Hussein, after
his capture in December 2003. DNA tests were carried out to prove the suspect was not one
of the many alleged “look alikes” of the former leader.

Firstly, the DNA was extracted from the mouth of the captured man using a swab. Great care
5 was taken to check that the swab did not become contaminated with any other DNA. DNA
extracted from the swab was then subjected to a standard technique called the polymerase
chain reaction (PCR), which takes a couple of hours. Lastly, the sample was “typed” to give
the genetic fingerprint. This was produced within 24 hours of Saddam Hussein’s capture.
Tests for use in criminal cases often take much longer because samples are very small or
10 contaminated.

It appears that Hussein’s genetic fingerprint was already stored away for comparison. This was
obtained from personal items such as his toothbrush. DNA from the toothbrush would have
been subjected to PCR before a DNA fingerprint could have been obtained.

Source: adapted from SHAONI BHATTACHARYA, New Scientist 15 December, 2003

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Use information from the passage and your own knowledge to answer the questions.

(a) Describe how the technique of genetic fingerprinting is carried out and explain how
it can be used to identify a person, such as Saddam Hussein.

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(6)

(b) Explain how DNA could be present on a toothbrush (line 12).

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(2)

(c) (i) Explain why the polymerase chain reaction was used on the sample of DNA from the
toothbrush (lines 12-13).

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(2)

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 (ii) Explain one way in which the polymerase chain reaction differs from DNA replication
in a cell.

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.............................................................................................................

.............................................................................................................

.............................................................................................................
(2)

(d) Tests for use in criminal cases often take much longer because samples are very small or
contaminated (lines 8-10). Explain why it takes longer to obtain a genetic fingerprint if the
sample is

(i) very small;

..............................................................................................................

.............................................................................................................
(1)

(ii) contaminated.

.............................................................................................................

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(2)
(Total 15 marks)

A gene was broken into fragments using enzyme Z. The mixture of fragments produced was
23 then separated by electrophoresis.

(a) What type of enzyme is enzyme Z?

......................................................................................................................
(1)

Page 39 of 92
 The table shows the number of base pairs present in the fragments.

Fragment Number of base pairs (× 103)

1 4.65

2 5.72

3 10.71

4 2.39

5 5.35

6 7.53

The diagram shows the electrophoresis gel used. The mixture of fragments was placed at the
start point marked S and the process started. The boxes indicate the positions reached by the
different fragments.

(b) Explain why base pairs are a suitable way of measuring the length of a piece of DNA.

......................................................................................................................

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(2)

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 (c) (i) Write 6 above the appropriate box on the diagram to show the position you would
expect fragment 6 to have reached.
(1)

(ii) Explain how you arrived at your answer.

.............................................................................................................

.............................................................................................................
(1)

(d) Enzyme Z recognises a particular sequence of bases in the gene. How many times does
this sequence appear in the DNA of this gene?

......................................................................................................................
(1)
(Total 6 marks)

(a) What is meant by a gene?

24
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......................................................................................................................
(2)

The polymerase chain reaction (PCR) can be used to obtain many copies of a particular gene.

(b) Explain how the strands of DNA are separated during the PCR.

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(2)

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 (c) In a particular PCR, two different primers are added to the DNA.

(i) Why are primers required?

.............................................................................................................

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(1)

(ii) Suggest why two different primers are required.

.............................................................................................................

.............................................................................................................
(1)

(d) Starting with a single molecule of DNA, the polymerase chain reaction was allowed to go
through three complete cycles. How many molecules of DNA would be produced?

Answer .......................................
(1)
(Total 7 marks)

Read the following passage.

25
The giant panda is one of the rarest animals in the world and is considered to be on the
brink of extinction in the wild. Giant pandas have been kept and bred in zoos with the hope
that they could be released into the wild. One worry is that small populations, like those in
zoos, reduce the genetic variation needed to allow the species to
5 adapt to changing conditions. Unfortunately, pandas find it difficult to reproduce in captivity.
Fertilisation of the females is guaranteed only by insemination with semen from several
males. With so many potential fathers, the true paternity of the cubs is not clear. It is
important to identify the fathers to maintain genetic variation.

10 Panda faeces can be collected in the wild. The faeces contain DNA from the panda, from
the bamboo on which they feed and from bacteria. The DNA is subjected to the polymerase
chain reaction (PCR). The primers used attach only to the panda DNA. The resulting DNA
is subjected to genetic fingerprinting. This can help us to count the number of individuals in
the wild because it allows us to identify individual pandas.

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Use information in the passage and your own biological knowledge to answer the questions.

(a) Describe how genetic fingerprinting may be carried out on a sample of panda DNA.

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(6)

(b) (i) Explain how genetic fingerprinting allows scientists to identify the father of a particular
panda cub.

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(2)

(ii) When pandas are bred in zoos, it is important to ensure only unrelated pandas breed.
Suggest how genetic fingerprints might be used to do this.

.............................................................................................................

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(1)

(c) (i) Suggest why panda DNA is found in faeces. (line 10)

.............................................................................................................

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(1)

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 (ii) Explain why the PCR is carried out on the DNA from the faeces. (line 12)

.............................................................................................................

.............................................................................................................
(1)

(iii) Explain why the primers used in the PCR will bind to panda DNA, but not to DNA
from bacteria or bamboo. (line 12)

.............................................................................................................

.............................................................................................................

.............................................................................................................

.............................................................................................................
(2)

(d) DNA from wild pandas could also be obtained from blood samples. Suggest two
advantages of using faeces, rather than blood samples, to obtain DNA from pandas.

1 ...................................................................................................................

......................................................................................................................

2 ...................................................................................................................

......................................................................................................................
(2)
(Total 15 marks)

Page 44 of 92
 Some species of crop plant produce a substance called glycinebetaine (GB).
26
Scientists transferred the gene for GB into a species of crop plant that does not normally produce
GB. These genetically modified plants then produced GB.

The scientists grew large numbers of the same crop plant with and without the gene at different
temperatures. After 3 days, they found the increase in dry mass of the plants.

Figure 1 shows their results.

Figure 1

(a) Describe the effect on growth of transferring the gene for GB into this plant.

........................................................................................................................

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(2)

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(b) The scientists measured the rate of photosynthesis in plants that produce GB and plants
that do not produce GB at 25°C, 35°C and 45°C.

Figure 2 shows their results.

Figure 2

(i) The scientists concluded that the production of GB protects photosynthesis from
damage by high temperatures.

Use these data to support this conclusion.

...............................................................................................................

...............................................................................................................

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(1)

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 (ii) Use the data from Figure 2 for plants that do not produce GB to explain the effect of
temperature on changes in dry mass of the plants shown in Figure 1.

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(Extra space)........................................................................................

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(4)

Rubisco activase is an enzyme found in chloroplasts. It activates the light-independent reaction

of photosynthesis.

The scientists discovered that, as temperature increased from 25°C to 45°C, rubisco activase
began attaching to thylakoid membranes in chloroplasts and this stopped it working.

(c) Rubisco activase stops working when it attaches to a thylakoid.

Use your knowledge of protein structure to explain why.

........................................................................................................................

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(2)

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(d) The scientists investigated the effect of GB on attachment of rubisco activase to thylakoid
membranes at different temperatures.

Figure 3 shows their results.

Figure 3

Use information from Figure 2 and Figure 3 to suggest how GB protects the crop plant
from high temperatures.

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(e) The scientists’ hypothesis at the start of the investigation was that crop plants genetically
engineered to produce GB would become more resistant to high environmental
temperatures.
The scientists developed this hypothesis on the basis of previous research on crops that
are grown in hot climates.

Suggest how the scientists arrived at their hypothesis.

........................................................................................................................

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(2)
(Total 15 marks)

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 Plasmids can be used as vectors to insert lengths of foreign DNA into bacteria. The diagram
27 shows how this is achieved.

(a) Name enzyme E.

......................................................................................................................
(1)

(b) Cut plasmids and lengths of foreign DNA can join. What features of their ends allows them
to join?

......................................................................................................................

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(2)

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(c) Draw three different structures that could be formed by incubating cut plasmids and lengths
of foreign DNA with ligase. Use the spaces provided on the diagram.

(3)
(Total 6 marks)

Page 51 of 92
 Huntington’s disease is a genetic condition that leads to a loss in brain function. The gene
28 involved contains a section of DNA with many repeats of the base sequence CAG. The number
of these repeats determines whether or not an allele of this gene will cause Huntington’s disease.

• An allele with 40 or more CAG repeats will cause Huntington’s disease.

• An allele with 36 – 39 CAG repeats may cause Huntington’s disease.

• An allele with fewer than 36 CAG repeats will not cause Huntington’s disease.

The graph shows the age at which a sample of patients with Huntington’s disease first developed
symptoms and the number of CAG repeats in the allele causing Huntington’s disease in each
patient.

(a) (i) People can be tested to see whether they have an allele for this gene with more than
36 CAG repeats. Some doctors suggest that the results can be used to predict the
age at which someone will develop Huntington’s disease.

Use information in the graph to evaluate this suggestion.

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 (ii) Huntington’s disease is always fatal. Despite this, the allele is passed on in human
populations. Use information in the graph to suggest why.

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(2)

(b) Scientists took DNA samples from three people, J, K and L. They used the polymerase
chain reaction (PCR) to produce many copies of the piece of DNA containing the CAG
repeats obtained from each person. They separated the DNA fragments by gel
electrophoresis. A radioactively labelled probe was then used to detect the fragments. The
diagram shows the appearance of part of the gel after an X-ray was taken. The bands
show the DNA fragments that contain the CAG repeats.

(i) Only one of these people tested positive for Huntington’s disease. Which person was
this? Explain your answer.

Person ..................................................................................................

Explanation ...........................................................................................

...............................................................................................................

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(2)

Page 53 of 92
(ii) The diagram only shows part of the gel. Suggest how the scientists found the number
of CAG repeats in the bands shown on the gel.

...............................................................................................................

...............................................................................................................

...............................................................................................................
(1)

(iii) Two bands are usually seen for each person tested. Suggest why only one band was
seen for Person L.

...............................................................................................................

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...............................................................................................................
(1)
(Total 9 marks)

Page 54 of 92
 Haemophilia is a genetic condition in which blood fails to clot. Factor IX is a protein used to treat
29 haemophilia. Sheep can be genetically engineered to produce Factor IX in the milk produced by
their mammary glands. The diagram shows the stages involved in this process.

Stage 1

Stage 2

Stage 3

Stage 4

Stage 5

Page 55 of 92
 Stage 6

(a) Name the type of enzyme that is used to cut the gene for Factor IX from human DNA
(Stage 1) .

........................................................................................................................
(1)

(b) (i) The jellyfish gene attached to the human Factor IX gene (Stage 2) codes for a protein
that glows green under fluorescent light. Explain the purpose of attaching this gene.

...............................................................................................................

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(2)

(ii) The promoter DNA from sheep (Stage 3) causes transcription of genes coding for
proteins found in sheep milk.

Suggest the advantage of using this promoter DNA.

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(Extra space) ........................................................................................

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(2)

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 (c) Many attempts to produce transgenic animals have failed. Very few live births result from
the many embryos that are implanted.

(i) Suggest one reason why very few live births result from the many embryos that are
implanted.

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(Extra space) ........................................................................................

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(2)

(ii) It is important that scientists still report the results from failed attempts to produce
transgenic animals. Explain why.

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(2)
(Total 9 marks)

(a) Adrenaline binds to receptors in the plasma membranes of liver cells. Explain how this
30 causes the blood glucose concentration to increase.

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(Extra space) .................................................................................................

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(2)

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(b) Scientists made an artificial gene which codes for insulin. They put the gene into a virus
which was then injected into rats with type I diabetes. The virus was harmless to the rats
but carried the gene into the cells of the rats.

The treated rats produced insulin for up to 8 months and showed no side-effects. The
scientists measured the blood glucose concentrations of the rats at regular intervals. While
the rats were producing the insulin, their blood glucose concentrations were normal.

(i) The rats were not fed for at least 6 hours before their blood glucose concentration
was measured. Explain why.

...............................................................................................................

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(1)

(ii) The rats used in the investigation had type I diabetes. This form of gene therapy may
be less effective in treating rats that have type II diabetes. Explain why.

...............................................................................................................

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(1)

(iii) Research workers have suggested that treating diabetes in humans by this method of
gene therapy would be better than injecting insulin. Evaluate this suggestion.

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 (Extra space) ........................................................................................

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(4)
(Total 8 marks)

Scientists wanted to measure how much mRNA was transcribed from allele A of a gene in a
31 sample of cells. This gene exists in two forms, A and a.

The scientists isolated mRNA from the cells. They added an enzyme to mRNA to produce cDNA.

(a) Name the type of enzyme used to produce the cDNA.

........................................................................................................................
(1)

The scientists used the polymerase chain reaction (PCR) to produce copies of the cDNA. They
added a DNA probe for allele A to the cDNA copies. This DNA probe had a dye attached to it.
This dye glows with a green light only when the DNA probe is attached to its target cDNA.

(b) Explain why this DNA probe will only detect allele A.

........................................................................................................................

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(2)

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(c) The scientists used this method with cells from two people, H and G.
One person was homozygous, AA, and the other was heterozygous, Aa.
The scientists used the PCR and the DNA probe specific for allele A on the cDNA from
both people.

The figure shows the scientists’ results.

(i) Explain the curve for person H.

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(3)

(ii) Which person, H or G, was heterozygous, Aa? Explain your answer.

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(2)
(Total 8 marks)

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 A husband and wife wanted to know whether they were carriers of the mutated form of a gene.
32 This mutation is a deletion that causes a serious inherited genetic disorder in people who are
homozygous.

A geneticist took samples of DNA from the husband and the wife. He used a DNA probe to look
for the deletion mutation. The DNA probe was specific to a particular base sequence in an exon
in the gene. Exons are the coding sequences in a gene.

The geneticist compared the couple’s DNA with that of a person known not to carry this mutation.

The chart shows the geneticist’s results.

(a) The geneticist told the couple they were both carriers of the mutated gene.
Explain how he reached this conclusion.

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(b) The DNA probe the geneticist used was for an exon in the DNA, not an intron. Explain why.

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(Extra space)..................................................................................................

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(3)

(c) To make the DNA probe, the geneticist had to find the base sequence of the normal gene.
Once he had copies of the gene, what methods would he use to find the base sequence of
the gene?

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(2)
(Total 8 marks)

Page 62 of 92
 Some populations of flies are becoming resistant to insecticides intended to kill them.
33
Scientists developed a method for finding out whether a fly was carrying a recessive allele, r, that
gives resistance to an insecticide. The dominant allele, R, of this gene does not give resistance.

The scientists:
• crossed flies with genotype RR with flies with genotype rr
• obtained DNA samples from the parents and offspring
• used the same restriction endonuclease enzymes on each sample, to obtain DNA
fragments.

(a) Explain why the scientists used the same restriction endonuclease enzymes on each DNA
sample.

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(2)

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 The scientists added two different primers to each sample of DNA fragments for the
polymerase chain reaction (PCR).

• Primer A3 only binds to a 195 base-pair fragment from allele r.

• Primer A4 only binds to a 135 base-pair fragment from allele R.

The scientists separated the DNA fragments produced by the PCR on a gel where shorter
fragments move further in a given time.

Their results are shown in Figure 1.

Figure 1

(b) Explain why primer A3 and primer A4 only bind to specific DNA fragments.

........................................................................................................................

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(2)

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(c) Use all the information given to explain the results in Figure 1.

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[Extra space] ................................................................................................

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(3)

(d) The scientists wanted to know on which chromosome the gene with alleles R and r was
located. From the flies with genotype RR, they obtained cells that were in mitosis and
added a labelled DNA probe specific for allele R. They then looked at the cells under an
optical microscope.

Explain why they used cells that were in mitosis.

........................................................................................................................

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........................................................................................................................

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(2)

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(e) Another group of scientists thought that pesticide resistance in some flies was related to
increased activity of an enzyme called P450 monooxygenase (PM).
This enzyme breaks down insecticides.

The scientists obtained large numbers of resistant and non-resistant flies. They then set up
the following experiments.

• Non-resistant flies exposed to insecticide.

• Resistant flies exposed to insecticide.
• Resistant flies treated with an inhibitor of PM and then exposed to insecticide.

They then determined the percentage of flies that were dead at different times after being
exposed to insecticide.

Figure 2 shows their results.

Figure 2

(i) Explain why the scientists carried out the control experiment with the non-resistant
flies.

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Page 66 of 92
 (ii) The scientists concluded that the resistance of the flies to the insecticide is partly due
to increased activity of PM but other factors are also involved.

Explain how these data support this conclusion.

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[Extra space] .......................................................................................

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(4)
(Total 15 marks)

Mycobacterium tuberculosis causes tuberculosis. The DNA of M. tuberculosis contains a direct

34 repeat (DR) region. The DR region consists of 43 different, non-coding base sequences called
spacers. Each spacer is found in a specific place in the DR region.
In different strains of M. tuberculosis, some of these spacers have been lost.

(a) (i) The DR region consists of non-coding base sequences.

What is meant by a non-coding base sequence?

...................................................................................................................

...................................................................................................................
(1)

(ii) Name the process by which the base sequence of a spacer is lost from a DR region.

...................................................................................................................

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(1)

Page 67 of 92
 Scientists investigated the DR regions of different strains of M. tuberculosis. They produced
a DNA probe for each of the 43 spacer sequences. Each probe was:

• labelled with a fluorescent marker that gave off light if the probe attached to its
complementary spacer
• attached to a particular square on a slide.

They obtained samples of the DR region from each strain. These were cut into small
single-stranded DNA fragments. The fragments from each strain were added to a slide with
the DNA probes attached. The diagram below shows their results for one strain of M.
tuberculosis with 20 of the probes.

(b) The scientists cloned the DR region DNA in vitro before testing for the presence of spacers.

Give the name of the method they used to clone the DNA in vitro.

.............................................................................................................................
(1)

(c) Explain how the use of DNA probes produced the results in the diagram.

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(d) Doctors can use the method with DNA probes to identify the specific strain of M.
tuberculosis infecting a patient. This is very important when there is an outbreak of a
number of cases of tuberculosis in a city.

Suggest and explain why it is important to be able to identify the specific strain of M.
tuberculosis infecting a patient.

.............................................................................................................................

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(2)
(Total 8 marks)

Page 69 of 92
 Agrobacterium tumefaciens is a bacterium that is often used in recombinant DNA technology to
35 produce transformed plants that benefit humans.

A. tumefaciens contains a plasmid which can be used as a vector to transfer a desired gene into
plant cells. These plant cells may then develop into plants which produce the protein coded for
by the desired gene.

The diagram outlines this process.

(a) (i) In stage 1, an enzyme is used to cut open the plasmid.

Name the type of enzyme used to cut open the plasmid.

...................................................................................................................
(1)

(ii) In stage 1, another enzyme is used to insert the desired gene into the plasmid DNA.
Name the type of enzyme used to insert the gene into the plasmid.

...................................................................................................................
(1)

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(b) In stage 4, some plant cells had plasmid DNA only in their cytoplasm. In other plant cells,
the plasmid DNA had become inserted into plant DNA in the nucleus.

In stage 5, only cells with plasmid DNA inserted into the plant DNA in the nucleus grew into
plants where all the cells contained the desired gene.

Explain why some of the plants in stage 5 contained the desired gene in all of their cells
and others did not.

.............................................................................................................................

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(3)

(c) The desired gene in the diagram was from an insect. In stage 6, the plant containing this
gene was able to use it to synthesise an insect protein.

The plant is able to synthesise the insect protein. Explain why this is possible.

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(3)
(Total 8 marks)

Page 71 of 92
Mark schemes
(a) use restriction enzyme / endonuclease / named, e.g. Bam / Eco;
1 to cut DNA in specific place / base sequence;
2

(b) heat DNA to 90 – 95 °C;

strands separate;
add primers;
and nucleotides;
cool so that primers bind to DNA;
(DNA) polymerase forms new strands / joins nucleotides;
4 max

(c) (i) virus is inhaled / sprayed into the lungs;

gets into cells, inserting the healthy gene;
2

(ii) makes DNA from RNA

rather than other way round
1
[9]

(a) (i) plasmid;

2 1

(ii) the bacteria divide / grow, producing many copies of desired

gene / plasmid;
OR
the bacteria divide / grow to cover the agar;
1

(iii) plant tissue that has antibiotic resistance survives;

identifies plant tissue which has desired gene / plasmid;
2

(iv) to clone plants / produce genetically identical plants with

gene / characteristic;
and produce large numbers / quickly;
2

(b) (i) (one reasonable suggestion),

e.g. toxin present all the time;
save costs of buying / application of spray;
no spray drift onto other fields / insects;
1 max

(ii) (one reasonable suggestion),

e.g. killing of harmless / useful insects that feed on wild plants;
damage to food chains starting with wild plants;
1 max
[8]

(a) 1. (If injected into egg), gene gets into all / most of cells of silkworm;
3 2. So gets into cells that make silk.
2
Page 72 of 92
 (b) 1. Not all eggs will successfully take up the plasmid;
2. Silkworms that have taken up gene will glow.
2

(c) Promoter (region / gene).

1

(d) 1. So that protein can be harvested;

2. Fibres in other cells might cause harm.
2
[7]

(a) (cut out gene using an) endonuclease / restriction enzyme;

4 reference to specificity / recognition site;
sticky ends;
use the same enzyme to cut;
plasmid / virus / potato DNA;
fixed by ligase;
method of introducing vector e.g. micropipette / virus injects DNA /
remove plant cell wall;
6 max

(b) different genes are expressed;

producing different enzymes / proteins;
2
[8]

(a) isolate wanted gene / DNA from another organism / mRNA from cell / organism;
5 using restriction endonuclease / restriction enzyme / reverse transcriptase to get DNA and
produce sticky ends;
use ligase to join wanted gene to plasmid;
also include marker gene e.g. antibiotic resistance;
add plasmid to bacteria to grow (colonies)then (replica) plate onto medium where the
marker gene is expressed;
bacteria / colonies not killed have antibiotic resistance gene and (probably) the wanted
gene;
6

(b) (i) injection, rapid rise and fall;

virus, slower rise and longer in effective / harmful range;
capsule slowest rise, longest in effective / harmful range;
injection and virus give harmful concentrations but capsule does
not;
3 max

Page 73 of 92
 (ii) advantage e.g.:
substance never reaches harmful levels / no side effect / less likely to harm the
organism, longer relief from symptoms / less frequent treatment needed /
longer effective range / longer but without harmful side effects;
1 max

disadvantage e.g.:
takes longer to take effect;
1
[11]

(a) (i) to separate polynucleotide strands / form single strands;

6 1

(ii) not denatured (at 95°C);

1

(iii) for binding of primers / nucleotides (to DNA strands);

1

(b) (i) doubling (of DNA) each cycle;

but very low numbers to start with, so appears flat then
exponential growth;
2

(ii) suggestion; with explanation e.g.:

nucleotides being used up;

so less / nothing to make complementary chains;

primers used up;

so cannot start complementary chains;

enzymes losing activity / denatured;

so no polymerisation of complementary strands;
2 max
[7]

(a) to separate the two strands / break hydrogen bonds;

7 1

(b) (i) enables replication / sequencing to start (allow keeps strands

separate);
1

(ii) joins DNA nucleotides (not complementary bases);

1

(c) (i) 64;

1

(ii) replication of DNA from crime scene / tissue sample /

for DNA sequencing / gene cloning;
1

Page 74 of 92
 (d) (transcription uses) RNA polymerase;
RNA nucleotides / uracil;
one (template) strand / PCR both strands;
start / stop codons;
(accept enzyme separates strands)
2 max
[7]

(a) probe will attach (to mutant allele);

8 attaches to one DNA strand;
as a result of complementary base pairing;
radioactivity detected on film / X-ray / by autoradiography
(if mutant allele present);
4

(b) for
gene is only active in mammary cells / only affects milk / easy to
obtain product / product produced in large amounts / gene passed to
offspring;
1

against
long term effects not known / qualified reference to animal exploitation
e.g. use of embryos / effect of inserted gene on other sheep
tissues / genes;
1
[6]

(a) introduction of healthy gene / ‘replacement’ of defective gene;

9 1

(b) can enter cells / infect cells / inject DNA into cells;
targets specific cells;
replicates (in cells);
2

(c) reproductive cells / gamete cells do not contain ADA allele / gene;
1

(d) (i) to ‘prevent’ rejection / immune response;

1

(ii) T lymphocytes have a limited life span / die off / do not reproduce;
bone marrow provides continual supply of T lymphocytes /
(ADA) gene enzyme;
2
[7]

Page 75 of 92
 1 DNA heated to 90 to 95°C;
10 2 strands separate;
3 cooled / to temperature below 70°C
4 primers bind;
5 nucleotides attach;
6 by complementary base pairing;
7 temperature 70 - 75°C;
8 DNA polymerase joins nucleotides together;
9 cycle repeated;
6 max
[6]

(a) (i) restriction (endonuclease) enzyme;

11
cuts DNA at specific / restriction points / after specific base sequence;
2

(ii) PCR / polymerase chain reaction;

1

(b) isolated cells divide by mitosis;

can get many plants (producing toxin) / rapid production of
(toxin producing) plants;
all cells (in the new plant / clone) will produce the toxin;
3
[6]

(a) (i) transfer / carry genes from one organism to another / into
12 bacteria / cells;
1

(ii) cut open plasmid;

cut donor DNA, to remove gene / length of DNA;
cut donor DNA and plasmid with the same enzyme / enzyme
that cuts at the same base sequence;
sticky ends / (overhanging) ends with, single strand / bases exposed;
association / attachment / pairing of complementary strand;
2 max

(iii) annealing / splicing / backbones joined / phosphodiester bonds;

1

(b) (i) L and M;

1

(ii) fragments 64 and 36(kilobases obtained)

1
[6]

Page 76 of 92
 (a) Restriction (enzyme / endonuclease);
13 1

(b) Move towards anode / move because charged;

Different rates of movement related to charge / size;

2

(c) (i) Piece of DNA;

Single stranded;
Complementary to / binds to known base sequence / gene;
max 2

(ii) DNA invisible on gel / membrane;

Allows detection;
2
[7]

(a) Presence of resistant and non-resistant varieties / mutation produces resistant variety;
14 Resistant ones survive / non-resistant ones killed by treatment;
These will reproduce and produce more resistant parasites / pass on resistance allele;
3

(b) Likelihood of being infected (by strain resistant to both drugs) is less;
1/500 × 1/500/1/250 000;
Drug has longer effective life;
max 2

(c) (i) As comparison / to show that nothing else in the treatment was responsible;
1

(ii) Given injections of saline / injection without SPf66;

(otherwise) treated the same as experimental group;
2

(d) (i) 100%;

1

(ii) 10%;
1

(e) (i) Different lengths of DNA have different base sequences / cut at specific
sequence;
Results in different shape / different shape of active site;
Therefore (specific sequence) will only fit active site of enzyme;
3

(ii) Recognition sites contain only AT pairs;

Which would occur very frequently;
2
[15]

Page 77 of 92
 (a) (i) contains genes / nucleotides / sections of DNA / artificial
15 DNA from two species / 2 types of organisms;
1

(ii) carries gene / DNA (into the other organism / gene carrier);
1

(iii) expose cells to the fungus;

non-resistant ones die, resistant ones survive;
OR identify by adding marker gene / gene probe / (qualified)
marker probe; description of positive result
e.g. radioactivity / fluorescence / complementary base pairing;
2

(b) EITHER 1 cut desired gene (from DNA) of oat plant;

2 using restriction endonuclease / restriction enzyme;
OR 1 use mRNA from oat which will code for resistance;
2 and use reverse transcriptase to form desired DNA;
OR 1 make artificial DNA with correct sequence of bases;
2 using DNA polymerase;
3 cut plasmid open;
4 with (same) restriction endonuclease / restriction enzyme;
5 ref. sticky ends / unpaired bases attached;
6 use (DNA) ligase to join / ref. ligation;
7 return plasmid to (bacterial) cells;
8 use of Ca2+ / calcium salts / electric shock;
(if ref. to ‘insulin’ allow 5 max.)
max 6
[10]

(a) only small amounts obtained / PCR increases the amount / mass of DNA;
16 so enough DNA available for genetic fingerprinting;
2

(b) (i) to separate the two strands of the DNA /

to break the hydrogen bonds;
(Reject “unzip”)
1

(ii) short lengths / fragments of DNA / nucleotides /

single stranded DNA;
1

(iii) to mark beginning and / or ends of the part of DNA needed /

for attachment of enzymes or nucleotides / initiator /
keeps strands apart;
1

(iv) would not be denatured;

must be heated to 95 °C / must withstand high temps;
2

Page 78 of 92
 (c) 1 DNA extracted from sample;
2 DNA cut / hydrolysed into segments using restriction endonucleases;
3 must leave minisatellites / required core sequences intact;
4 DNA fragments separated using electrophoresis;
5 detail of process e.g. mixture put into wells on gel and electric
current passed through;
6 immerse gel in alkaline solution / two strands of DNA separated;
7 Southern blotting / cover with nylon / absorbent paper (to absorb DNA);
8 DNA fixed to nylon / membrane using uv light
9 radioactive marker / probe added (which is picked up by required
fragments) / complementary to minisatellites;
10 (areas with probe) identified using X-ray film / autoradiography;
max 6

(d) adult 3;
this is only one which, (with number 1), can provide (all) the DNA
fragments which children have / all bars match;
(Reject ‘genes’)
2
[15]

(a) (i) Sticky ends / description;

17 Reference to complementary base-pairing
2

(ii) Ligase;
1

(b) Carrier of DNA / gene; (context of foreign DNA)

Into cell / other organism / host;
2

(c) Act as marker gene;

Allows detection of cells containing plasmid / DNA;
2
[7]

(a) Mother and father both heterozygotes / Tt / carriers;

18 Probability of thalassaemia 1/4 and female 1/2;
Probability of both 1/8;
3

(b) (i) Cut at same base sequence as same enzyme used;

Fragments are same length / size / have same charge;
2

(ii) Single base occurs many times;

Sequence of 20 unlikely to occur elsewhere;
Allow one mark for establishing the principle where neither marking
point clearly made.

2
[7]

Page 79 of 92
 (a) (i) Sticky ends / description;
19 Reference to complementary base-pairing
2

(ii) Ligase;
1

(b) Carrier of DNA / gene; (context of foreign DNA)

Into cell / other organism / host;
2

(c) Act as marker gene;

Allows detection of cells containing plasmid / DNA;
2
[7]

(a) (i) Different genes / characteristics / features;

20 Reference to mutations;
Or
Base sequence determines protein;
Different species have different protein sequences;
max 2

(ii) Primer has different DNA sequence;

DNA specific / complementary base-pairing;
2

(iii) Electrophoresis separates DNA;

(So they can be) identified by position on gel;
Smaller / shortest fragments travel furthest / quicker / or
reverse argument;
3

(b) (conventional) Many lengths / all DNA / (new) one length;

Each rung is DNA of one / specific length;
2

(c) 1 Heat DNA;

2 Breaks hydrogen bonds / separates strands;
3 Add primers;
4 Add nucleotides;
5 Cool;
6 (to allow) binding of nucleotides / primers;
7 DNA polymerase;
8 Role of (DNA) polymerase;
9 Repeat cycle many times;
max 6
[15]

Page 80 of 92
 (a) (i) Reverse transcriptase;
21 1

(ii) Idea that mRNA is present in large amounts in cell making

the protein / mRNA has been edited / does not contain
introns / mRNA codes for single protein;
1

(b) (Ligase) splices / joins two pieces of DNA / “sticky ends”;

1
[3]

(a) 1. DNA is cut;

22 2. using restriction enzyme;
3. electrophoresis;
4. separates according to length / mass / size;
5. DNA made single-stranded;
6. transfer to membrane / Southern blotting;
7. apply probe;
8. radioactive / single stranded / detected on film / fluorescent;
9. reference to tandem repeats / VNTRs / minisatellites;
10. pattern unique to every individual;
6 max

(b) cells on toothbrush;

DNA present in cell;
2

(c) (i) toothbrush gives small sample of DNA / need more DNA
for analysis;
PCR gives many copies;
2

(ii) uses heat;

to separate strands;
OR
PCR replicates pieces of DNA;
because DNA has been cut;
OR
primer added in PCR;
to initiate replication
2 max

(d) (i) PCR / amplification needed;

1

(ii) other DNA present; need to identify ‘required’ DNA from rest;
2
[15]

Page 81 of 92
 (a) Endonuclease / restriction enzyme;
23 1

(b) DNA made of base pairs;

Each base pair is same length / occupies same distance
along backbone;
2

(c) (i) Second blank box from left labelled 6;

1

(ii) Distance moved depends on length / number of base pairs /

second longest fragment / second shortest distance identified;
1

(d) 5;
1
[6]

(a) a length of DNA;

24 that codes for a single protein / polypeptide;
2

(b) by heating;
to break the H-bonds (between complementary bases);
2

(c) (i) to allow the DNA polymerase to attach / start addition of

nucleotides / mark start and end of sequence to be
copied / prevents strands re-joining;
1

(ii) because the sequences at the ends of the target sequence

are different / one is at the beginning and one at the end;
1

(d) 8;
accept 7
1
[7]

Page 82 of 92
 (a) 1 DNA is cut;
25
2 Using restriction enzyme;

3 Use electrophoresis;

4 Separates according to length / mass;

5 Southern blotting / transfer to (nylon) membrane;

6 Make single-stranded;

7 Apply probe;

8 Radioactive / fluorescent;

9 Reference to tandem repeats / VNTRs / minisatellites;

10 Autoradiography / eq;
8 and 10 should be consistent
max 6

(b) (i) All bands in cub which don’t come from mother;

Must be in father’s DNA fingerprint;

Principle that all bands in cub must come from mother and father =
1
2

(ii) Select pairs with dissimilar DNA fingerprints;

1

(c) (i) Cells (from panda) in faeces / gut cells / blood cells;
1

(ii) To increase amount of DNA / only small amount present;

1

(iii) DNA / primer has specific base-sequence;

Reference to specific / complementary base-pairing;
2

(d) Taking samples from animals causes stress / injury to animal;

Difficult to find animals;

Pandas are dangerous / threat to human;

max 2
[15]

Page 83 of 92
 (a) 1. No effect at 25°C
26
The question only refers to plants with GB
1. Reject same mass

2. Keeps growing at 30°C and 35°C / up to 35°C (more than without GB);

3. Above 35°C, falls but grows more than plant without GB;
3. Accept at all temperatures above 25°C more growth than without
GB
2 max

(b) (i) Significantly different / SEs do not overlap ;

Accept converse without GB
1

(ii) (As temperature increases,)

1. Enzyme activity reduced / (some) enzymes denatured;

2. Less photosynthesis, so fewer sugars formed;

3. Less respiration / less energy / ATP for growth;

4. Less energy for named function associated with growth

4. Eg mitosis, uptake of mineral ions
4

(c) 1. (Rubisco activase attaches to thylakoid and) this changes shape / tertiary
structure (of enzyme) / blocks active site / changes active site;
Note - question states enzyme stops working when it attaches to
thylakoid, not before
1. Accept rubisco in this context

2. (This) prevents substrate / RuBP entering active site / binding;

2. Accept prevents ES complex forming
2. Accept no longer complementary to substrate / RuBP
2

(d) 1. GB prevents / reduces binding of rubiscoactivase to (thylakoid membrane);

1. Accept enzyme instead of rubiscoactivase. Accept rubisco

2. (Prevents it) up to 35°C;

3. (So) rubiscoactivase / enzyme remains active;

4. (So) photosynthesis / light-independent stage still happens;

4. Accept descriptions of light-independent stage

5. Above 35°C, some binding still occurs but less than without GB, so less
reduction in growth;
4 max

Page 84 of 92
 (e) 1. Looked for information / journals, on crop plants that grow at high temperatures;
1. “other research” is minimum accepted
1. Accept previous experiments research with temperature resistant
crops
Ignore simple references to looking at previous studies / other
plants - need to relate to this context

2. (Crop plants cited in this research) contain / make GB;

3. So assumed making plants produce GB makes them resistant to high

temperatures;
2 max
[15]

(a) restriction (enzyme) / endonuclease / named example;

27 1

(b) unpaired bases / sticky ends / staggered;

complementary / explained;
2

(c) 1 mark for each correct outcome

plasmid with foreign DNA joined in ring;
ring with plasmid only; ring of foreign DNA only;
ignore linear structures
3
[6]

(a) (i) 1. Negative correlation;

28
Accept: description for ‘negative correlation’
Neutral: ‘correlation’
Reject: positive correlation

2. Wide range;

3. Overlap;

4. (Graph suggests that) other factors may be involved (in age of onset);
2 / 3 Accept the use of figures from the graph
2 / 3 Can refer to age of onset or number of CAG repeats
Ignore references to methodology
3 max

Page 85 of 92
 (ii) 1. Age of onset can be high / symptoms appear later in life;
Accept: ‘gene’ for ‘allele’

2. (So) individuals have already had children / allele has been passed on;

OR

3. Individuals have passed on the allele / already had children;

4. Before symptoms occur;

2 max

(b) (i) 1. Person K;

2. (As has) high(est) band / band that travelled a short(est) distance / (er) so
has large(st) fragment / number of CAG repeats;
Must correctly link distance moved and fragment size
2

(ii) Run fragments of known length / CAG repeats (at the same time);
Accept: references to a DNA ladder / DNA markers
Do not accept DNA sequencing
1

(iii) Homozygous / (CAG) fragments are the same length / size / mass;
Accept: small fragment has run off gel / travelled further
1
[9]

(a) Restriction / endonuclease;

29
Ignore specific names of restriction enzymes e.g. EcoR1
1

(b) (i) 1. (Acts as a) marker gene to show that the (human) gene has been taken
up / expressed;
1. Accept: gene marker

2. (Only) implant cells / embryos that show fluorescence / contain the

jellyfish gene;
2

(ii) 1. Factor IX present in / extracted from milk;

2. Gene only expressed in mammary glands / udder / gene not expressed

elsewhere;
2. Ignore references to milk
The ‘only’ aspect is important here.

3. Do not need to kill sheep (to obtain Factor IX);

2 max

Page 86 of 92
 (c) (i) 1. Mutation / nucleus / chromosomes / DNA may be damaged / disrupts
genes;
1. Neutral: cell may be damaged

2. May interfere with proteins (produced) / gene expression / translation;

Ignore references to hormone levels or time of implantation

OR

3. Embryo / antigens foreign;

3. Neutral: antigens change

4. Embryo is rejected / attacked by immune system;

4. sNeed idea that the immune system is involved if mark point 3
has not been given
‘Embryo foreign so rejected’ = 2 marks
‘Embryo rejected by immune system’ = 1 mark
‘Embryo is rejected’ = 0 marks
2 max

(ii) 1. Saves time / money for others;

2. Same work is not repeated / methods can be compared / improved /

amended / same errors are not made;
2
[9]

(a) 1. Adenylate cyclase activated / cAMP produced / second messenger produced;

30
2. Activates enzyme(s) (in cell so) glycogenolysis / gluconeogenesis occurs /
glycogenesis inhibited;
2. Neutral: ‘glucose produced’ as given in the question stem
Accept: correct descriptions of these terms
2

(b) (i) 1. Glucose / sugar in food would affect the results;

1. Accept references to starch / carbohydrate
Or

2. Food / eating would affect blood glucose (level);

Or

3. (Allows time for) blood glucose (level) to return to normal;

3. Neutral: allows time for insulin to act
1 max

(ii) Type 2 diabetes is a failure to respond to insulin / still produces insulin / is not
insulin-dependent;
1

Page 87 of 92
 (iii) (For) – 3 max
A maximum of three marks can be awarded for each side of the
argument

1. Avoids injections / pain of injections;

2. Long(er) lasting / permanent / (new) cells will contain / express gene;

Ignore references to methodology e.g. sample size not known

3. Less need to measure blood sugar / avoids the highs and lows in blood
sugar;

4. Less restriction on diet;

(Against) – 3 max

5. Rats are different to humans;

6. May have side effects on humans;

6. Accept: virus may be harmful / disrupt genes / cause cancer

7. Long(er) term effects (of treatment) not known / may have caused effects
after 8 months;

8. (Substitute) insulin may be rejected by the body;

4 max
[8]

(a) Reverse transcriptase;

31 1

(b) 1. Probe (base sequence) complementary (to DNA of allele A / where A is (and)
binds by forming base pairs / hydrogen bonds;
Accept gene A

2. So (only) this DNA labelled / has green dye / gives out (green) light;
Accept glows for green light
2

(c) (i) 1. More probe binding / more cDNA / mRNA / more allele / gene A means
more light;

2. DNA (with A) doubles each (PCR) cycle;

3. So light (approximately) doubles / curve steepens more and more (each

cycle) / curve goes up exponentially / increases even faster;
3

Page 88 of 92
 (ii) (G because)

1. (Heterozygous) only has half the amount of probe for A attaching / only
half the amount of DNA / allele A (to bind to);
Accept only one A to bind to

2. (So,) only produced (about) half the light / glow / intensity (of H) (per cycle
of PCR);
If reference to ‘half’ for point 1, allow ‘less light’ in 2.
2
[8]

(a) 1. Carriers are heterozygous / have one normal copy and one mutant copy of gene /
32 have one recessive allele / don't have the condition;

2. Both have DNA that binds (about) half / 50% amount of probe (that non-carrier
does);

3. Probe binds to dominant / healthy allele so only one copy of exon in their DNA /
have one copy of gene without exon / base sequence for probe to bind to;
3. Accept normal and gene
3. Accept have a deletion mutation
3

(b) 1. Introns not translated / not in mRNA / (exons) code for amino acids / introns do
not code for amino acids;
1. Accept not expressed
1. Accept polypeptide / protein for amino acids

2. Mutations of these (exons) affect amino acid sequences (that produce) faulty
protein / change tertiary structure of protein;
2. Accept deletion leads to frameshift
2. In this context, accept affects protein made

3. So important to know if parents’ exons affected, rather than any other part of
DNA / introns;
Accept converse arguments involving - eg introns do not code for
amino acids / proteins
Reject references to making amino acids, once
3

(c) 1. Restriction mapping / described;

2. DNA / base sequencing (of fragments) / description / name of method;

2
[8]

Page 89 of 92
 (a) 1. Cut (DNA) at same (base) sequence / (recognition) sequence;
33
Accept: cut DNA at same place

2. (So) get (fragments with gene) R / required gene.

Accept: ‘allele’ for ‘gene’ / same gene
2

(b) 1. Each has / they have a specific base sequence;

2. That is complementary (to allele r or R).
Accept description of ‘complementary’
2

(c) 1. Fragments L from parent rr, because all longer fragments / 195
base pair fragments;
Ignore: references to fragments that move further / less, require
identification of longer / shorter or 195 / 135
Accept: (homozygous) recessive

2. Fragments N from parent RR, because all shorter fragments / 135 base pair
fragments;
1 and 2 Accept: A3 for 195 and A4 for 135
2. Accept: (homozygous) dominant

3. (M from) offspring heterozygous / Rr / have both 195 and 135 base pair
fragments.
Accept: have both bands / strips
Reject: primer longer / shorter
3

(d) 1. (Cells in mitosis) chromosomes visible;

2. (So) can see which chromosome DNA probe attached to.
2

(e) (i) 1. For comparison with resistant flies / other (two) experiments
/ groups;
Ignore: compare results / data / no other factors

2. To see death rate (in non-resistant) / to see effect of insecticide in

non-resistant / normal flies.
Accept: ‘pesticide’ as ‘insecticide’
Accept to see that insecticide worked / to see effect of enzyme
2

Page 90 of 92
 (ii) (PM must be involved because)
1. Few resistant flies die (without inhibitor);
2. More inhibited flies die than resistant flies;
3. (PM) inhibited flies die faster (than resistant flies);
(Other factors must be involved because)
4. Some resistant flies die;
5. But (with inhibitor) still have greater resistance / die slower than
non-resistant flies.
Accept: (with inhibitor) die slower than non-resistant flies
4 max
[15]

(a) (i) Does not code for amino acid/tRNA/rRNA;

34
Accept ‘does not code for production of protein/polypeptide’
Reject ‘that produces/makes amino acid'
1

(ii) Deletion mutation;

Accept ‘deletion’
Ignore references to splicing
1

(b) (The) polymerase chain reaction;

Accept PCR
1

(c) 1. Probes are single stranded / have a specific base sequence;

2. Complementary base sequence on (specific) spacer

OR

3. Complementary/specific to (particular) spacer;

4. (In white squares probe) binds (to single-stranded spacer) and
glows/produces light/fluoresce;
2. Need idea of complementary to spacer
3. Accept converse for dark squares
3

Page 91 of 92
 (d) 1. To see if strain is resistant to any antibiotics;
2. So can prescribe effective/right antibiotic;

OR

3. To see whether (any) vaccine works against this strain/ see

which vaccine to use/ to produce specific vaccine;
4. (So) can vaccinate potential contacts/to stop spread;

OR

5. Can test other people to see if they have the same strain/ to
trace where people caught TB;
6. Allowing control of spread of disease/vaccinate/treat contacts
(of people with same strain) before they get TB;
Do not allow mix and match of points from different alternative pairs
2 max
[8]

(a) (i) Restriction endonuclease;

35 1

(ii) (DNA) ligase;

1

(b) (For those plants that contained the desired gene in the
nucleus/plant DNA)

1. (DNA of desired gene) copied/replicated with host DNA/inside

nucleus;
2. Passed on by mitosis/plant grows by mitosis;
3. Produces genetically identical cells/clones;
Ignore references to protein synthesis or plasmids not taking up the
gene
1. Accept DNA replication during mitosis
1. and 2. Accept converse for plants with the gene in the cytoplasm
3. Neutral ‘identical unqualified’
3. Accept description, e.g., DNA is the same
3

(c) 1. Genetic code is universal/triplets in DNA always code for

same amino acid;
2. It/insect DNA can be transcribed;
3. Can be translated (process/mechanism same in all
organisms/cells);
2. Accept (basic) transcription (process/mechanism) same in all
organisms/cells;
2. Accept descriptions of process
3. Accept descriptions of process
3
[8]

Page 92 of 92