H

RAFFLES INSTITUTION
Higher 2

CANDIDATE NAME

CIVICS
GROUP

S0

INDEX
NUMBER

1 2

BIOLOGY

9648/03
18 th September 2012
2 hours

Paper 3
Additional materials:

Answer Booklet/Paper

READ THESE INSTRUCTIONS FIRST

Write your index number, CT group and name on all the work you hand in.
Write in dark blue or black pen on both sides of the paper.
You may use a soft pencil for any diagrams, graphs or rough working.
Do not use staples, paper clips, highlighters, glue or correction fluid.
Answer all questions.
At the end of the examination, hand in your essay SEPARATELY.
The number of marks is given in brackets [ ] at the end of each question
or part question.

For Examiners Use

1
2
3
4
5
Total

This document consists of 16 printed pages.

Answer all the questions in the spaces provided.Raffles Institution

Internal Examination

Restriction enzymes cut DNA molecules only at specific target sites with particular
base sequences. The target sites for enzymes EcoRI, BamHI and EcoRV are
RI 2012

Preliminary Examination 9648/03

2
shown in Table 1.1. The lines in each sequence show where the enzyme cuts the
DNA molecule.
Restriction enzyme

Specific target base sequence of DNA

G A A T T C

EcoRI

C T T A A G

BamHI

EcoRV

G G A T C C
C C T A G G
G A T A T C
C T A T A G
Table 1.1

(a)

Some restriction enzymes generate sticky ends, while others generate blunt ends.
Explain why it is more efficient to clone DNA fragments with sticky ends than DNA
fragments with blunt ends.
....
.......[1]

The pcDNA plasmid shown in Fig.1.1 is a very useful cloning vector used in
recombinant DNA technology. In the commercial production of insulin, the cDNA
of human insulin is inserted into pcDNA.
The recombinant pcDNA is then transformed into Escherichia coli to produce
[Turn over
RI 2012

Preliminary Examination 9648/03

For
Examiners
Use

eI
Nh e I
Pm a I
Ap a I
Xb o I
Xh t I
No XI
Bst RV
Eco RI
Eco I
X
Bst H I
m
Ba n I
Kp d III
Hin

large numbers of recombinant plasmids. The recombinant plasmids are then

purified and transfected into mammalian cells which will express high levels of the
functional insulin.

Fig.1.1
(b)

A genetic engineer cuts the plasmid shown in Fig.1.1 with EcoRV in order to
insert a human insulin gene.
Describe the steps that must be carried out after digestion with EcoRV to produce
a recombinant plasmid.
....
....
....
....
....
....
.......[4]

(c)

The pcDNA contains two selectable markers, the ampicillin resistance gene
(AmpR) and the neomycin resistance gene (NeoR) which code for resistance to the
antibiotics ampicillin and neomycin respectively. Ampicillin specifically targets cell
wall synthesis whilst neomycin binds irreversibly to ribosomes preventing protein
synthesis. The pcDNA used has been modified to express the NeoR only in the
[Turn over

RI 2012

Preliminary Examination 9648/03

For
Examiners
Use

4
For
Examiners
Use

mammalian cells.
(i)

Explain why the AmpR is not an effective selectable marker in mammalian cells.
....
....
....
....
.......[2]

(ii)

Suggest why the NeoR is only expressed in mammalian cells.

....
.......[1]

(d)

Explain why mammalian cells instead of E.coli were used to produce the
functional human insulin.
....
....
....
....
.......[2]
[Total: 10]

Osteogenesis imperfecta (brittle bone

characterised by bones that break easily.

disorder),

is

genetic

disorder

Adult bone stem cells transplanted into children with osteogenesis imperfecta
have stimulated growth of bone in these children. During the 6 months
RI 2012

Preliminary Examination 9648/03

5
immediately following the transplant, the childrens growth reached 60% to 94% of
expected normal values for children their age.
(a)

(i)

In terms of differentiation potential, explain how an adult bone stem cell is

different from a fertilized egg of a human.
....
....
....
.......[2]

(ii)

Explain how the adult bone stem cells may increase bone growth in children with
osteogenesis imperfecta.
....
....
....
.......[2]

(b)

The polymerase chain reaction (PCR) can be used to make many copies of a
gene. Fig.2.1 shows the main stages in the process.

Stage 1:
Stage 3: DNA is heated
2: DNA
cooled
to 64oC
DNA is heated to 95oC for 5 minutes in the presence of primers, DNA Stage
nucleotides
& is
DNA
polymerase

the cycle is repeated

Fig.2.1
(i)

Describe the unique property of the DNA polymerase used in stage 1.

(ii)

.......[1]
Identify stage 3 and state the temperature required at this stage.
.......[1]

(iii)

Explain the role of the primers in Fig.2.1.

[Turn over

RI 2012

Preliminary Examination 9648/03

For
Examiners
Use

....
....
....
.......[3]
(iv)

How many cycles of the PCR would be needed to produce 128 molecules of DNA
from a single DNA molecule?
.......[1]

(c)

The sequence of nucleotides in a sample of DNA can be determined using a

method involving the PCR followed by electrophoresis. In this method, the
primers used in the PCR are radioactively labelled and some of the normal
deoxyribonucleotides are chemically altered.
Fig.2.2 shows a single strand of DNA and the copies produced by the PCR using
a mixture of all 4 DNA nucleotides (dATP, dTTP, dGTP and dCTP) and some
chemically altered dTTP also known as ddTTP. This ddTTP is represented as T*
in Fig.2.2.

T*

T*
T*
T*

Fig.2.2
(i)

In order for this method to work, both electrophoresis and the use of radioactive
primers are necessary. Explain why this is so.
....
....
.......[2]
Fig.2.3 shows the chemical structure of deoxyribonucleotide (left) and a
chemically altered deoxyribonucleotide (right).
.

RI 2012

Preliminary Examination 9648/03

For
Examiners
Use

7
For
Examiners
Use

Fig.2.3
(ii)

With reference to the Figs.2.2 and 2.3, explain why three different lengths of new
strand were produced.
....
....
....
.......[2]

(d)

Scientists have found a new method of copying DNA that is faster than PCR. The
new method is called Helicase Dependent Amplification (HDA). HDA uses an
enzyme to separate the two strands of DNA.
This means that DNA can be copied at a constant temperature of 37C. In all
other ways, HDA works in exactly the same way as PCR.
HDA is faster than PCR. Explain why.
....
.......[1]
[Total: 15]

(a)

Plant physiologists attempted to produce papaya plants using tissue culture.

They investigated the effects of different concentrations of two plant growth
factors on small pieces of the stem tip from a papaya plant. Their results are
[Turn over

RI 2012

Preliminary Examination 9648/03

8
For
Examiners
Use

shown in the Table 3.1.

Table 3.1
(i)

With reference to the Table 3.1, give evidence that the cells from the stem tip are
totipotent.
....
....
....
.......[2]

(ii)

Stem tips are used in tissue culture to produce disease-free plants.

Explain why cells are taken from the stem tip instead of flowers or leaves for
tissue culture.
....
....
....
....
....

(iii)

.......[2]
State the recommended cytokinin to auxin ratio that should be used to produce
papaya plants using stem tip culture.
.......[1]

RI 2012

Preliminary Examination 9648/03

9
For
Examiners
Use

(iv)

Explain the advantage of growing papaya plants from tissue culture rather than
from seeds.
....
....
....
....
....
.......[3]

(b)

In 1989, a transgenic salmon line was created by injecting eggs of the Atlantic
salmon with a gene construct containing the gene of the Chinook salmon growth
hormone. Transgenic salmon were grown as sterile, all-female populations in
[Turn over

RI 2012

Preliminary Examination 9648/03

10
tanks on land located away from the sea.
Salmon reaches a marketable value at 4kg. However, this line of salmon was not
allowed to be commercialized. The growth of the transgenic salmon and the
standard Atlantic salmon is shown in Figure 3.1.

(i)

Fig.3.1
With reference to Fig.3.1, explain the benefits of farming transgenic salmon as
compared to the standard salmon.
....
....
....
.......[3]

(ii)

RI 2012

Give reasons why transgenic salmon are grown as sterile, all-female populations
in tanks on land away from the sea.

Preliminary Examination 9648/03

For
Examiners
Use

11
....
....
....
.......[2]
The gene construct used is shown in Fig.3.2. It contains a promoter and
termination region from the ocean pout antifreeze gene and a growth hormone
from a Chinook salmon.

Fig.3.2
(iii)

Explain why the promoter from the ocean pout antifreeze gene is used in this
construct as opposed to the original growth promoter of the Atlantic salmon.
....
....
....
.......[2]
[Total:15]

Planning question
You are required to plan, but not carry out, an investigation into the effect of substrate
(hydrogen peroxide, H2O2) concentration on the rate of enzymatic activity of yeast cells.
[Turn over
RI 2012

Preliminary Examination 9648/03

For
Examiners
Use

12

Yeast cells contains enzyme catalase which catalyses the decomposition of H2O2.
H2O2 H2O + O2
When droplets of a yeast-alginate mixture are dropped into calcium chloride solution,
yeast-alginate beads are produced and the yeast is immobilized into alginate beads.
When the one of the yeast-alginate beads is then transferred to a test-tube containing
H2O2, it will naturally settle at the bottom of the tube. Over time oxygen bubbles will form
around the bead. The bubbles provide buoyancy and will cause the bead at the bottom of
the test-tube to float to the top.
The rate of catalase activity can be determined by measuring the time taken for the yeastalginate bead to float.
Your planning must be based on the assumption that you have been provided with the
following equipment and materials, which you must use:
1M hydrogen peroxide
Yeast-alginate mixture
Calcium chloride solution
Droppers
Forceps
Test-tube
A variety of different sized beakers, measuring cylinders or syringes for measuring
volumes.
Stop watch
Hot water (80oC)
Your plan should:
have a clear and helpful structure such that the method you use is able to be
repeated by anyone reading it,
be illustrated by relevant diagram(s) to show, for example, the arrangement of the
apparatus used, if necessary.
include layout of results tables and graphs with clear headings and labels,
include full details and explanations of the procedures that you would adopt to
ensure that the results obtained were as quantitative, precise and reliable as
possible,
include relevant risks and precautions taken,
the correct use of technical and scientific terms.
[Total: 12]

RI 2012

Preliminary Examination 9648/03

For
Examiners
Use

13
For
Examiners
Use

....
....
....
....
....
....
....
....
....
....
....
....
....
....
....
....
[Turn over
RI 2012

Preliminary Examination 9648/03

14

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

RI 2012

Preliminary Examination 9648/03

For
Examiners
Use

15
....
....
....
....
....
....
....
....
....
....
....
....
....
....
....
....
....
....
....
....
....
....
....
....
....
Free-response question
[Turn over
RI 2012

Preliminary Examination 9648/03

For
Examiners
Use

16
For
Examiners
Use

Write your answer to this question on the separate paper provided.

Your answer :
should be illustrated by large, clearly labelled diagrams, where appropriate
must be in continuous prose, where appropriate.
must be set out in section (a), (b)., as indicated in the question.
5

(a)

Describe the genetic basis of cystic fibrosis and, compare liposomal and viral delivery
systems in the treatment of cystic fibrosis using gene therapy.
[8]

(b)

Discuss how the human genome project can help parents "customise" their babies
and the possible social and ethical implications of doing so.
[6]

(c)

Describe how RFLP can be used in detecting sickle-cell anaemia in a human

population.
[6]
[Total:20]

RI 2012

Preliminary Examination 9648/03