New Senior Secondary Mastering Biology (Second Edition) E4: Chapter 1

Question Bank Multiple-choice questions

% Chapter 1 Techniques in modern

biotechnology

Multiple-choice questions

[10171289]
* The base sequences of two alleles of a gene are shown below. One of them is the
normal allele and the other is the mutated allele.

By cutting the two alleles with the same restriction enzyme separately and
looking at the numbers of fragments produced, scientists can differentiate the
two alleles. Which of the following shows the recognition site of a restriction
enzyme that can be used by the scientists to differentiate the two alleles?
A AAGCTT
B GAATCC
C GAGCTC
D GGATCC
C
---------------------------------------------------

[10171293]
* During polymerase chain reaction, the reaction mixture is heated and cooled
repeatedly. Which of the following is the reason for cooling the reaction
mixture?
A To prevent the denaturation of DNA polymerase.
B To allow the annealing of primers to the single-stranded DNA.
C To separate the double-stranded DNA into two single strands.
D To provide a suitable temperature for the extension of primers by joining
free nucleotides that are complementary to the templates.
B
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New Senior Secondary Mastering Biology (Second Edition) E4: Chapter 1
Question Bank Multiple-choice questions

[10171295]
**The diagram below shows two alleles of a gene, one of them is the normal allele
and the other is the mutated allele. Some of the base sequences of the alleles are
shown.

Different numbers of fragments are produced after cutting the two alleles
separately with a restriction enzyme having a recognition site CCGG. If the
resulting fragments are separated by gel electrophoresis, which of the following
shows the pattern of bands on the gel?
A B

C D

D
---------------------------------------------------

[10171308]
* Which of the following statements about cloning animals using nuclear transfer
is correct?
A The animals produced are called genetically modified animals.
B The animals produced have the same genetic information as their ovum
donors.
C The animals produced may look different from their ovum donors.
D The animals produced must have more desirable characters than their
nuclear donors.
C
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New Senior Secondary Mastering Biology (Second Edition) E4: Chapter 1
Question Bank Multiple-choice questions

[10171323]
 The table below shows the functions of two enzymes that are commonly used in
genetic engineering.
Enzyme Function
catalyses the addition of nucleotides to a
X
DNA strand
catalyses the formation of covalent bonds
Y
between two DNA fragments

Which of the following correctly identifies X and Y?

X Y
A DNA polymerase restriction enzyme
B DNA polymerase DNA ligase
C DNA ligase DNA polymerase
D restriction enzyme DNA ligase
B
---------------------------------------------------

[10171324]
* Directions: The following two questions refer to the steps involved in restriction
fragment length polymorphism (RFLP) analysis shown below.
(1) Add radioactive DNA probes.
(2) Cut DNA samples using restriction enzymes.
(3) Denature DNA into single strands.
(4) Separate DNA fragments by gel electrophoresis.

Which of the following is the correct order of these steps?

A (2) → (3) → (4) → (1)
B (2) → (4) → (3) → (1)
C (3) → (1) → (2) → (4)
D (3) → (2) → (1) → (4)
B
---------------------------------------------------

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New Senior Secondary Mastering Biology (Second Edition) E4: Chapter 1
Question Bank Multiple-choice questions

[10171330]
* The pattern of bands obtained in RFLP analysis is unique to each individual
because
A DNA probes bind to variable number tandem repeats (VNTRs) at different
loci in different individuals.
B the lengths of VNTRs are different in different individuals.
C the amount of DNA is different in different individuals.
D the restriction enzymes recognize different base sequences in different
individuals.
B
---------------------------------------------------

[10171366]
* Directions: The following two questions refer to the steps involved in the
production of pest-resistant plants shown below.
(1) Infect plant cells with bacteria.
(2) Insert the toxin gene into plasmids.
(3) Obtain plasmids from bacteria.
(4) Introduce plasmids into bacteria.

Which of the following is the correct order of these steps?

A (1) → (4) → (2) → (3)
B (3) → (2) → (4) → (1)
C (3) → (4) → (2) → (1)
D (4) → (2) → (3) → (1)
B
---------------------------------------------------

[10171368]
* Which of the following bacteria is used in steps (1), (3) and (4)?
A Escherichia coli
B Vibrio cholerae
C Staphylococcus aureus
D Agrobacterium
D
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New Senior Secondary Mastering Biology (Second Edition) E4: Chapter 1
Question Bank Multiple-choice questions

[10171371]
* Directions: The following two questions refer to the diagram below, which
shows the major steps in recombinant DNA technology.

In which steps are a restriction enzyme and a DNA ligase used?

Restriction enzyme DNA ligase
A step 1 step 3
B step 2 step 1
C step 2 step 3
D step 3 step 2
C
---------------------------------------------------

[10171375]
* Why does the plasmid used in recombinant DNA technology carry an antibiotic
resistance gene?
A It allows the selection of the recombinant plasmids.
B It allows the selection of the transformed host cells.
C It helps the host cells pick up the recombinant plasmids.
D It kills the host cells which are not transformed.
B
---------------------------------------------------

[10171378]
 Tissue culture is a method of
A introducing a foreign gene into an organism.
B producing useful products in an organism.
C producing genetically identical organisms.
D producing organisms with new characteristics.
C
---------------------------------------------------

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New Senior Secondary Mastering Biology (Second Edition) E4: Chapter 1
Question Bank Multiple-choice questions

[10171392]
* Directions: The following two questions refer to the diagram below, which
shows the basic steps involved in the production of genetically modified plants.

Restriction enzymes are used in

A step 1 only.
B step 2 only.
C steps 2 and 3 only.
D steps 3 and 4 only.
C
---------------------------------------------------

[10171397]
* Why is Agrobacterium used in the above process?
(1) Agrobacterium contains plasmids.
(2) Agrobacterium can infect plant cells.
(3) Part of the plasmids of Agrobacterium can integrate into DNA of plant cells.
A (1) and (2) only
B (1) and (3) only
C (2) and (3) only
D (1), (2) and (3)
D
---------------------------------------------------

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New Senior Secondary Mastering Biology (Second Edition) E4: Chapter 1
Question Bank Short questions

Short questions

[10171535]
* Gel electrophoresis can be used to separate pieces of DNA on the basis of their
molecular size.
Small DNA fragments travel further along the electrophoresis gel than large
fragments.
 The restriction enzyme MstII recognizes the sequence CCTNAGG – where
N can be any DNA nucleotide.
 Sickle-cell anaemia is caused by a mutation in the β-globin gene.
 The mutation changes CCTGAGG to CCTGTGG.
 Mutated DNA produces longer fragments of DNA.
a Using MstII as an example, explain what is meant by a restriction enzyme.
(3 marks)
b Explain why longer fragments of DNA occur in the mutated form of the β-
globin gene. (2 marks)
c Samples of DNA from three individuals were treated with the restriction
enzyme MstII and then loaded onto an electrophoresis gel.
 Individual X is homozygous for normal haemoglobin and has the
normal β-globin gene.
 Individual Y is homozygous for sickle-cell anaemia and has the
mutated form of the β-globin gene.
 Individual Z is heterozygous.
Following electrophoresis, the DNA was located using a radioactive probe.
The diagram below shows the electrophoresis gel and the relative positions
of the DNA fragments for individuals X and Y.

Complete the diagram by drawing the position of the DNA from individual

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New Senior Secondary Mastering Biology (Second Edition) E4: Chapter 1
Question Bank Short questions

Z in the blank column on the diagram. (1 mark)

OCR GCE (A) Human Biology Genetics, Control and Ageing Jun 2012 Q2a–c

-- answer --
a A restriction enzyme is an enzyme that cuts DNA strands at specific sites. 1m
For example, MstII cuts only at the sites with base sequence CCTNAGG. 1m
Some restriction enzymes produce sticky ends and some produce blunt ends.
1m
b There are fewer recognition sites in the mutated form of the β-globin gene. 1m
The restriction enzyme cuts at fewer sites. 1m
c One band in line with the band of X and one band in line with the band of Y 1m
---------------------------------------------------

[10171539]
* Look at the diagram.
It shows the cloning technique used to produce Dolly the sheep.

a Describe the cloning technique used to produce Dolly.

Use the diagram to help you. (3 marks)
b Is Dolly a clone of sheep X or sheep Y?
Explain your answer. (1 mark)
c Some people object to the cloning of mammals.
Suggest why. (1 mark)
OCR GCSE Biology B Higher Tier Unit 1 Jun 2008 Q10
-- answer --

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Question Bank Short questions

a The nucleus of an ovum from sheep X is removed. 1m

The nucleus of a body cell from sheep Y is placed into the ovum. 1m
The ovum is then put into the surrogate sheep. 1m
b Dolly is a clone of sheep Y because the DNA / genes / chromosomes of Dolly
came from sheep Y. 1m
c Some people object to the cloning of mammals due to religious or ethical
reasons. /
Some people think that money could be better spent on new medicines. /
Some people object to the cloning of mammals because cloning reduces the
variations among individuals of the same species. /
Some people object to the cloning of mammals because the animal clones might
have abnormalities or problems like premature ageing. 1m
---------------------------------------------------

[10171548]
* The flow chart below shows steps in the process of plant tissue culture.

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New Senior Secondary Mastering Biology (Second Edition) E4: Chapter 1
Question Bank Short questions

a Outline the general purpose of plant tissue culture. (1 mark)

b Sterile conditions are used throughout the process. Explain why sterile
conditions are necessary. (1 mark)
c Describe how callus tissue may be stimulated to initiate root and shoot
development. (1 mark)
d State two advantages of producing plants by plant tissue culture rather than
by seeds. (2 marks)
e Plant tissue culture is increasingly used in conjunction with genetic
engineering to propagate transgenic plants. Outline two additional steps,
further to the steps shown above, which would be required in the production
of transgenic plants. (2 marks)
CCEA GCE (A) Biology Module 5 Jan 2010 Q2

-- answer --
a To produce large numbers of genetically identical plants / clones. 1m
b To prevent the growth of fungi and bacteria, which would utilize the nutrients in the
culture medium and damage the plant tissue. 1m
c Add hormones to the culture medium. 1m
d Very large numbers of plants can be produced from a single explant. /
All plants produced are genetically identical. /
All plants produced are disease-free. /
Plants (such as orchids) that are hard to produce from seeds can be propagated.
(any 2) 1m x 2
e Use of a vector to introduce the gene of interest into plant cells. 1m
Selection of the plant cells containing the gene of interest. 1m
---------------------------------------------------

[10171552]
**The enzyme, polygalacturonase (PG), is responsible for the softening of fruits
such as tomatoes. To prolong the shelf life of tomatoes, scientists have
genetically engineered tomato plants to create the ‘Flavr Savr’ tomato that does
not bruise during shipping and lasts longer on the shelf.
To do this, the PG gene responsible for the production of the enzyme was first
isolated. Scientists then used this isolated gene to create an ‘anti’ or ‘nonsense’
PG gene that was then introduced into tomato plants. This nonsense gene stops
the translation of the original PG gene in the tomato plant and so stops enzyme
production.

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New Senior Secondary Mastering Biology (Second Edition) E4: Chapter 1
Question Bank Short questions

The diagram below summarizes the action of the introduced nonsense PG gene.

a Suggest how the nonsense PG gene could have been introduced into the
tomato plant. (3 marks)
b Name the process labelled X in the diagram. (1 mark)
c Suggest two ways by which matching of the sense and nonsense mRNA
would prevent translation and so inhibit the production of the
polygalacturonase enzyme. (2 marks)
CCEA GCE (AS) Biology Module 1 Jun 2008 Q7b–d

-- answer --
a The nonsense PG gene and a bacterial plasmid are cut with the same restriction
enzyme. /
The nonsense PG gene is inserted into the plasmid using a DNA ligase. /
The recombinant plasmid is taken up by Agrobacterium. /
Transformed Agrobacterium is used to infect tomato plant tissue.
(any 3) 1m x 3

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New Senior Secondary Mastering Biology (Second Edition) E4: Chapter 1
Question Bank Short questions

b Transcription 1m
c The double-stranded mRNA cannot exit the nucleus. /
The double-stranded mRNA cannot bind with ribosomes. /
The codons on the mRNA are not available for the binding of anticodons on the
tRNA. (any 2) 1m x 2
---------------------------------------------------

[10171574]
* The diagram below shows how carrot plants can be cloned by tissue culture.

a Is tissue culture a form of sexual reproduction or asexual reproduction?

Explain your answer. (1 mark)
b Suggest one advantage of producing plants by tissue culture. (1 mark)
c Explain why a batch of plants produced by tissue culture is more easily
destroyed by viral infection than a batch of plants grown from seeds.
(1 mark)
d Apart from nutrients and oxygen, name one other substance that must be
added into the culture medium. Explain why this substance is needed.
(2 marks)
-- answer --
a It is a form of asexual reproduction because no fusion of gametes is involved.
1m
b Desirable characters of selected plants can be preserved. /
It can be used as a method to overcome reproductive difficulties of plants. 1m
c It is because plants produced by tissue culture are genetically identical / lack
genetic variations. 1m
d Hormones 1m
They are used to promote the growth of roots and shoots. 1m
---------------------------------------------------

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New Senior Secondary Mastering Biology (Second Edition) E4: Chapter 1
Question Bank Short questions

[10171578]
 Complete the following paragraph with suitable words. (3 marks)
A DNA probe is a short, a -stranded DNA fragment with a known b
. It can be used to detect DNA fragments containing a sequence which is c
to its sequence.
-- answer --
a single 1m
b base sequence 1m
c complementary 1m
---------------------------------------------------

[10171584]
 Complete the following paragraph with suitable words. (4 marks)
a is a method to clone animals. The first mammal cloned by this
method was Dolly the sheep. This method can be used to propagate b
animals to help maintain biodiversity. However, the clones produced lack genetic
variations. This reduces the
c of the population to changes in the environment. Cloning can also
be used to create embryos, which can provide
d for research and medical treatment.

-- answer --
a Nuclear transfer 1m
b endangered 1m
c adaptability 1m
d stem cells 1m
---------------------------------------------------

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New Senior Secondary Mastering Biology (Second Edition) E4: Chapter 1
Question Bank Structured questions

Structured questions

[10171760]
* Tissue culture is a method of artificial propagation in plants. One technique for
carrying out tissue culture is to remove meristems from shoot tips of plants and
culture them as shown in the diagram below.

a State one advantage of using meristems in plant tissue culture. (1 mark)

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New Senior Secondary Mastering Biology (Second Edition) E4: Chapter 1
Question Bank Structured questions

b With reference to the above diagram, explain how a large number of

plantlets are produced from one meristem. (3 marks)
c Outline two advantages and two disadvantages of using this technique
instead of seed production to produce food plants. (4 marks)
OCR GCE (A) Growth, Development and Reproduction Jun 2010 Q3a–c

-- answer --
a Meristems are rapidly dividing. 1m
b Meristematic cells divide by mitotic cell division to form a mass of cells. /
The cells are separated. /
Each cell contains genes that are required to produce a new plant. /
The cells differentiate. /
Each cell grows into a plant. (any 3) 1m x 3
c Advantages:
As all the plants produced are identical, plants with desirable characters can be
produced. /
It is a rapid way to mass produce plants. /
It can be used to produce plants from genetically modified plant cells. /
It can be used to produce plants that are hard to grow from seeds.
(any 2 or other reasonable answers) 1m x 2
Disadvantages:
The process is labour intensive. /
Highly trained staff is required to carry out the process. /
Producing plants using this method is expensive. /
Sterile conditions are required when the process is carried out. /
The plants produced lack genetic variations.
(any 2 or other reasonable answers) 1m x 2
---------------------------------------------------

[10171772]
* The human protein, insulin, is involved in the homeostatic control of blood
sugar. Most artificially produced human insulin is currently produced by
genetically modified bacteria, but several companies are now investigating the
possibility of producing human insulin using the safflower plant, a plant which is
normally grown for its oil. This method would allow large scale production and,
in theory, one large North American farm would be capable of meeting the global
demand for insulin.
The first stage in this process would be the isolation and removal of the human

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New Senior Secondary Mastering Biology (Second Edition) E4: Chapter 1
Question Bank Structured questions

insulin gene. Once isolated, the gene is then placed into a vector. One example of
such a vector is the bacterium, Agrobacterium tumefaciens, a relatively common
soil bacterium that regularly infects plants.
a Describe how the human insulin gene could be inserted into the bacterium,
Agrobacterium tumefaciens. (4 marks)
The genetically modified bacterium is then allowed to infect a sample of cells
extracted from the safflower plant and these infected cells can be grown into
mature plants, each being capable of producing human insulin. The human
insulin produced by these plants appears to be fully effective in the treatment of
diabetes mellitus. However, a patient’s blood insulin cannot be raised by simply
eating transgenic safflower plants.
b Suggest why eating safflower plants containing insulin would not result in
an increase in the patient’s blood insulin levels. (2 marks)
One possible risk of growing a genetically modified crop in this way is that it
might interbreed with wild plants. Pharmaceutical companies try to minimize
such risk by growing the crop ‘counter-seasonally’ to reduce the chances of the
insulin gene being transferred to other plants.
c Suggest what is meant by the term ‘counter-seasonally’. (1 mark)
CCEA GCE (AS) Biology Module 1 Jun 2009 Q7b–d

-- answer --
a A plasmid obtained from the bacterium and the human insulin gene are cut with
the same restriction enzyme.
1m
The sticky ends allow the plasmid and the human insulin gene to bind together
with hydrogen bonds. 1m
A DNA ligase catalyses the joining of the plasmid and the human insulin gene by
helping the formation of covalent bonds. 1m
A bacterium takes up the plasmid. 1m
b Insulin is a protein. It will be digested / broken down if it is ingested. 1m
The absorbed amino acids cannot be reassembled into insulin in the body due to
the lack of a functional gene. 1m
c Genetically modified plants flower when other plants are not flowering to reduce
the chances of cross-pollination. 1m
---------------------------------------------------

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Question Bank Structured questions

[10171776]
* The diagram below represents the processes involved in the polymerase chain
reaction (PCR). This technique allows forensic scientists to analyse and
potentially match DNA samples left at the scene of a crime.

a What advantage does PCR offer to forensic scientists? (1 mark)

b i In step 1, what is the purpose of heating the DNA sample to 95 °C?
(2 marks)
ii Primers, each containing roughly twenty bases are also added to the
DNA sample in step 1. What is the purpose of these primers in the
process? (1 mark)
iii Name the enzyme that would be added to the DNA sample in step 2.
(1 mark)
iv What else should be added to the process in step 2 to allow successful
replication of the DNA sample? (1 mark)
c In step 3, replication of the DNA is completed. Describe the sequence of
events, following the separation of the DNA strands in step 1, which lead to
the DNA being replicated. (3 marks)

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New Senior Secondary Mastering Biology (Second Edition) E4: Chapter 1
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The heating of the DNA sample to 95 °C in step 1 could denature the enzymes
involved.
d i Describe precisely how high temperatures could denature the enzymes.
(1 mark)
ii Suggest how scientists have solved this potential problem. (1 mark)
e In some legal cases, the use of PCR has been discredited. Suggest one
possible source of error in the PCR process. (1 mark)
CCEA GCE (AS) Biology (New Specification) Module 1 Jun 2009 Q7

-- answer --
a A large number of DNA copies can be synthesized from a single DNA sample. /
It can be used in DNA fingerprinting. 1m
b i Heating breaks the hydrogen bonds between DNA strands. 1m
Two separate strands are formed. / Both strands can act as templates.
1m
ii Primers stop the two DNA strands from rejoining. /
Primers initiate the action of DNA polymerase. /
Primers can be used to bracket the regions of DNA to be amplified.
1m
iii DNA polymerase 1m
iv Free nucleotides 1m
c The mixture is cooled to allow the binding of primers with the strands of DNA. /
Temperature is then increased. /
Each DNA strand acts as a template. /
Nucleotides bind to the complementary bases on DNA (A–T, C–G, T–A, G–C). /
Nucleotides are held in place by hydrogen bonds between the complementary
bases. /
Adjacent nucleotides are joined by forming covalent bonds. (any 3) 1m x 3
d i Heating breaks the bonds that hold the enzymes in shape / changes the
active sites of the enzymes. 1m
ii Use heat-stable enzymes / enzymes extracted from thermophilic bacteria.
1m
e The DNA sample may be contaminated. /
Errors may occur in the replication process.
(or other reasonable answers) 1m
---------------------------------------------------

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Question Bank Structured questions

[10171788]
* a Nucleic acids are composed of nucleotides. Complete the table below to
give two ways in which the structure of a DNA nucleotide differs from that
of an RNA nucleotide.
DNA nucleotide RNA nucleotide
1
2
(2 marks)
b The polymerase chain reaction (PCR) enables many copies of DNA to be
made from a small sample. The diagram below summarizes the procedure.

i Explain what is happening at each of the steps indicated. (3 marks)

ii At the end of the first cycle, there are two molecules of DNA.
How many molecules will there be at the end of five cycles?
(1 mark)
c Explain, precisely, how restriction enzymes cut DNA molecules into
fragments. (2 marks)
d A DNA fragment, consisting of 24 base pairs, was analysed for the number
of different bases on each strand. The table below shows some of the
results. Determine the missing values and complete the table.
Number of bases
A G T C
Strand 1 7 8
Strand 2 4
(2 marks)

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New Senior Secondary Mastering Biology (Second Edition) E4: Chapter 1
Question Bank Structured questions

CCEA GCE (AS) Biology (New Specification) Module 1 Jan 2009 Q7

-- answer --
a
DNA nucleotide RNA nucleotide
A DNA nucleotide has the sugar A RNA nucleotide has the sugar
1
deoxyribose. ribose.
A DNA nucleotide may have the A RNA nucleotide may have the
2
base thymine. base uracil.
1m x 2
b i Step 1:
Heat the reaction mixture to separate the DNA strands / break the hydrogen
bonds to release the two strands. 1m
Step 2:
Add primers to the reaction mixture. 1m
Step 3:
Use DNA polymerase to extend the primers by joining free nucleotides which
are complementary to the templates. / Nucleotides attach to the DNA
strands through complementary base pairing (A pairs with T and C pairs with
G). 1m
ii 32 1m
c Restriction enzymes cut DNA molecules at specific base sequences, 1m
across the sugar-phosphate backbone. 1m
d
Number of bases
A G T C
Strand 1 7 8 4 5
Strand 2 4 5 7 8
2m
---------------------------------------------------

[10171789]
**A restriction enzyme, EcoRI, acts on DNA by cutting it at the sequence shown
below.

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New Senior Secondary Mastering Biology (Second Edition) E4: Chapter 1
Question Bank Structured questions

a What is a sticky end? (1 mark)

b State the base sequence of the bases in the sticky end on strand Y.
(1 mark)
c The diagram below shows the positions of four restriction sites, W, X, Y and
Z, in a plasmid. The distances between these sites are measured in kb (1 kb
= 1000 base pairs)

Scientists mixed EcoRI with the plasmid and he separated the resulting
DNA fragments by gel electrophoresis. The diagram below shows the
positions of the DNA bands in the gel.

It is known that two of the restriction sites in the plasmids can be cut by
EcoRI. Which of the restriction sites were cut? Explain your answer.
(4 marks)
d Explain why the DNA fragments moved at different speed in the gel.
(1 mark)

-- answer --
a A sticky end is a short length of single-stranded DNA at the cut end of the DNA

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New Senior Secondary Mastering Biology (Second Edition) E4: Chapter 1
Question Bank Structured questions

fragments. 1m
b TTAA 1m
c X and Z 2m
The 60 kb DNA fragment consists of the 10 kb and 50 kb segments. 1m
The 40 kb DNA fragment consists of the 15 kb and 25 kb segments. 1m
d The molecular spaces in the gel slab allowed shorter DNA fragments to move
faster than the longer ones. 1m
---------------------------------------------------

[10171790]
* Human insulin can be produced using genetic engineering. The diagram below
shows part of the process.

a Name two enzymes that are used in stage X. (2 marks)

b Many copies of the DNA encoding human insulin are required. Name the
technique which can be used to amplify a specific DNA fragment from a
small amount of DNA. (1 mark)
c i In stage X, which structure in the bacterial cell is the human DNA
incorporated into? (1 mark)
ii Explain the role of this structure in genetic engineering. (2 marks)
iii Describe three properties of this structure which make it ideal for use
in genetic engineering. (3 marks)

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New Senior Secondary Mastering Biology (Second Edition) E4: Chapter 1
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d Explain why the bacteria are grown in culture medium containing an

antibiotic. (2 marks)
e In the past, insulin for injection was extracted from the pancreases of cattle
or sheep. State two advantages of producing insulin using genetic
engineering over the extraction of insulin from animal pancreases.
(2 marks)
-- answer --
a Restriction enzyme 1m
DNA ligase 1m
b Polymerase chain reaction 1m
c i Plasmid 1m
ii The plasmid acts as a vector 1m
to carry the DNA encoding human insulin into the bacterial cell. 1m
iii It can pick up foreign DNA easily. /
It can be picked up by host cells because of its small size. /
It can replicate independently of the host cell’s chromosome and exist in
multiple copies in host cells. /
It usually carries an antibiotic resistance gene which allows the selection of
transformed cells. (any 3) 1m x 3
d To select the transformed bacteria. 1m
Only the transformed bacteria which carry an antibiotic resistance gene survive in
the culture medium. 1m
e The insulin produced is not rejected by the immune system. /
The insulin produced is pure. /
The product yield is much higher. /
The extraction cost is lower. (any 2) 1m x 2
---------------------------------------------------

[10171791]
* Scientists tried to produce a genetically modified bacterium, which can be used
to make a useful protein. They first obtained some DNA fragments containing
the gene for the protein and performed polymerase chain reaction (PCR). The
flow chart below shows the steps in one PCR cycle.

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New Senior Secondary Mastering Biology (Second Edition) E4: Chapter 1
Question Bank Structured questions

a Explain why PCR was performed. (1 mark)

b Why was the reaction mixture heated to 95 °C? (1 mark)
c What is the role of the primers? (1 mark)
d Explain why heat-stable DNA polymerase was used in PCR. (1 mark)
e If one DNA molecule was present in the reaction mixture before PCR
started, how many copies of the DNA molecules were present in the
reaction mixture after five PCR cycles? (1 mark)
f After cycles of PCR, the DNA fragments containing the gene for the protein
were cut using a restriction enzyme. Three restriction enzymes and their
corresponding recognition sites are shown below.
Restriction enzyme Recognition site

i Explain why X and Y are more useful than Z in recombinant DNA

technology. (2 marks)
ii The base sequence of the DNA fragment containing the gene for the
protein is shown below.

The scientists want to insert the gene into plasmids. Which restriction
enzymes, X or Y, should be used to cut the DNA fragments? Explain
your answer. (3 marks)
iii Name an enzyme that can be used to join the resulting DNA fragments
and the plasmids together. (1 mark)
g The plasmids used in recombinant DNA technology usually carry an
antibiotic resistance gene. What is the importance of this? (1

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New Senior Secondary Mastering Biology (Second Edition) E4: Chapter 1
Question Bank Structured questions

mark)
-- answer --
a To amplify the gene. /
To increase the quantity of the gene. 1m
b To separate the double helix of the DNA fragment into two single DNA strands by
breaking the hydrogen bonds between the base pairs in the DNA. 1m
c To bracket the DNA region to be amplified. /
To serve as starting points for DNA synthesis. /
To stop the two DNA strands from rejoining. 1m
d Heat-stable DNA polymerase would not be denatured when the reaction mixture
was heated to high temperatures in PCR cycles. 1m
e 25 = 32 copies 1m
f i X and Y leave sticky ends on the DNA fragments after cutting. 1m
The unpaired bases in the sticky ends of DNA fragments cut with the same
restriction enzyme can bind together with hydrogen bonds. This facilitates
the joining of the DNA fragments.
1m
ii X 1m
X cuts outside the gene. 1m
Y cuts within the gene. This destroys the gene. 1m
iii DNA ligase 1m
g The antibiotic resistance gene acts as a selective marker to allow the selection of
transformed cells. 1m
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[10171796]
* Genetically modified bacteria can be used to produce human insulin. DNA
encoding human insulin is introduced into bacteria by recombinant DNA
technology. In the process, a restriction enzyme is used to cut DNA.
BamHI is a commonly used restriction enzyme. Its recognition site is shown
below.

a The base sequence of a DNA fragment is shown below. Draw dotted lines to
shown how BamHI cuts the DNA fragment. (2 marks)

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New Senior Secondary Mastering Biology (Second Edition) E4: Chapter 1
Question Bank Structured questions

b Explain why BamHI is useful in recombinant DNA technology. (2 marks)

c Name the enzyme used to join DNA fragments. (1 mark)
A recombinant plasmid is shown below.

The recombinant plasmid is mixed with bacteria. The bacteria are then cultured
on an agar plate containing the antibiotic ampicillin.
d Explain why the bacteria have to be cultured on an agar plate containing
ampicillin. (3 marks)
The bacteria on the agar plate are then cultured in fermenters. They produce
polypeptides, which can be processed into human insulin.
e State two advantages of producing insulin using this method over the
extraction of insulin from animal pancreases. (2 marks)
-- answer --
a

1m x 2
b BamHI leaves sticky ends on the DNA fragments after cutting. 1m
The unpaired bases in the sticky ends of the DNA fragments cut with the same
restriction enzyme can bind together with hydrogen bonds. This facilitates the
joining of the DNA fragments. 1m
c DNA ligase 1m
d Only some of the bacteria have picked up the recombinant plasmid / are
transformed. 1m
Only the transformed bacteria which carry an ampicillin resistance gene survive
on the agar plate containing ampicillin. 1m
Culturing the bacteria on the agar plate containing ampicillin allows the selection
of transformed bacteria. 1m
e The insulin produced is not rejected by the immune system. /

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New Senior Secondary Mastering Biology (Second Edition) E4: Chapter 1
Question Bank Structured questions

The insulin produced is pure. /

The product yield is much higher. /
The extraction cost is lower. (any 2) 1m x 2
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New Senior Secondary Mastering Biology (Second Edition) E4: Chapter 1
Question Bank Structured questions

[10171802]
**Read the following article and answer the questions.
Leukaemia is a kind of cancer. Scientists found that in some patients it is
caused by a specific type of translocation mutation.
Polymerase chain reaction (PCR) can be used to detect this mutation for
the diagnosis of leukaemia. mRNA is first extracted from the cells of a person
who is suspected to have leukaemia. Single-stranded DNA is synthesized using
mRNA.

The DNA is then amplified by PCR. Two different primers are used. Each
primer can bind to a region on the DNA or its complementary strand of DNA.
The two regions flank the junction where two chromosome segments join in
the translocation mutation.

The DNA fragments in the PCR product are then separated by gel
electrophoresis. A specific band will appear in the gel only if the person has

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New Senior Secondary Mastering Biology (Second Edition) E4: Chapter 1
Question Bank Structured questions

the mutation.

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New Senior Secondary Mastering Biology (Second Edition) E4: Chapter 1
Question Bank Structured questions

a What is a translocation mutation? (1 mark)

b State how mutations may lead to the onset of cancer. (1 mark)
c Calculate how many double-stranded DNA molecules would be present in
the reaction mixture at the end of cycle 6 of PCR. (2 marks)
d Explain why a specific band will appear in the gel only if the person has the
translocation mutation in the genome. (3 marks)
e Suggest two advantages of using PCR in disease diagnosis. (2 marks)

-- answer --
a A translocation mutation is the exchange of segments of chromosomes between
non-homologous chromosomes. 1m
b Mutations in the DNA that controls cell division may lead to uncontrollable division
of the cells. 1m
c 2(6–1) 1m
= 32 1m
d The translocation mutation brings the segments of chromosomes from two
chromosomes together. 1m
If the mutation does occur, the two primers will bind to both the DNA strands in
cycle 2 and amplification of the DNA molecule results. 1m
If the mutation does not occur, only one primer will bind to one of the two DNA
strands in cycle 2. Amplification of the DNA molecule cannot be carried out. 1m
e It is a fast method. 1m
Only a small amount of sample is needed. 1m
(or other reasonable answers)
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