HKALE BIOLOGY | 29 Applied Genetics | P.

Recombinant DNA technology

1. 1999/IIB/7
Genetic engineering is the artificial manipulation of the genome of a organism. It includes gene cloning
and / or clonal propagation. Gene cloning refers to the creation of a recombinant organism. Clonal
propagation is the mass production of the recombinant organism.
State three different fields in which gene cloning and clonal propagation can be applied. Use specific
examples to illustrate the applications in each field. Outline the procedure used in one of the examples you
cited.
Concern has been raised regarding the safety and bioethics of genetic engineering. State one concern
about each of these two aspects.
(20 marks)
2. 2000/IIB/8
Genetically modified (GM) foods contain components from GM organisms. Transgenic technology is one
major way to produce GM organisms. Outline the principles of using transgenic technology to make GM
organisms for producing GM foods. In what ways can the yield and quality of food be enhanced by this
technology? Suggest some measures to minimize the potential risks of this new technology to our health
and environment.
(20 marks)
3. 2006/IIA/2ci
In some people, a gene mutation results in the formation of a defective digestive enzyme that may cause a
gastrointestinal disease.
One way to treat this disease is by enzyme replacement, i.e. introducing an external source of the digestive
enzyme into the patients’ gastrointestinal tract. The enzyme used can be produced by recombinant DNA
technology. Outline how the enzyme is produced by this technology. (4 marks)

3. 2007/IC/12
(c) The gene coding for the toxic protein was originally found in the bacteria Bacillus thuringiensis (also
known as Bt). Briefly outline the principles of the technology involved in the transfer of this Bt gene
from the bacteria into the cultured cells of cotton plants. (lines 11 –15)
(4 marks)
(d) The production of Bt cotton is a means of crop protection against insect pests. Describe two
advantages of using this approach over the use of insecticides. (2 marks)

DNA fingerprinting
1. 2004/IB/11
Three DNA samples were used in the following analysis. These included linear viral DNA, plasmid DNA
and DNA marker. The same amount of DNA from each sample was completely cut with a restriction
enzyme. This was followed by gel electrophoresis where the DNA fragments moved from the negative to
the positive pole. The gel was then stained with a dye so that bands containing DNA fragments could be
visualized under ultra-violet light.
The DNA marker has a known molecular size in basepair (bp). The marker served as a standard to
measure the molecular size of DNA fragments. A photograph of the electrophoresis pattern is shown
below:
 HKALE BIOLOGY | 29 Applied Genetics | P.2

(a) Based on the pattern shown, determine the number of sites that can be cut by the restriction enzyme
in the
(i) plasmid DNA (1 mark)
(ii) linear viral DNA (1 mark)
(b) Based on the pattern shown, the molecular size of the DNA marker is 28 000 bp (found by the sum of
the molecular size of its DNA fragments). What is the molecular size of the plasmid DNA? Show your
calculation.
(N.B. Assume that the molecular size of an individual DNA fragment is measured by the leading edge
of the fragment facing the positive pole.)
(1 mark)
(c) According to the pattern shown, explain whether the viral DNA or the plasmid DNA has a larger
molecular size. (2 marks)
(d) Suppose the patterns of DNA fragments shown are unique to the virus and the plasmid,
(i) what is the scientific name for these patterns? (1 mark)
(ii) state two applications of these viral and plasmid DNA patterns. (2 marks)
(e) If the same amount of human DNA is completely cut by the above-mentioned restriction enzyme and
electrophoresed in the same way, a smear with no discrete DNA bands is obtained. Account for this
result. (3 marks)

2. 2005/IA/8
DNA fingerprinting can be applied in different areas.
(a) One application of DNA fingerprinting is to identify the relationship among members in a family. The
result of a DNA fingerprinting test performed on four individuals, P, Q, R and S, is shown below:

Based on the data above, deduce with reasons the most probable relationship of the following persons with
P:
(i) Q (2 marks)
(ii) S (2 marks)
(b) Another application of DNA fingerprinting is to identify the evolutionary relationship among different
groups of living organisms. What is the assumption involved in this application of DNA fingerprinting?
(1 mark)

3. 2007/IIA/3d
Referring to postulate IV, modern genetic technology can be used to study whether the re-isolated
microorganism may be similar to the suspected one. Name this technology and outline the principles
involved. (5 marks)
 HKALE BIOLOGY | 29 Applied Genetics | P.3

Gene therapy
1. 2006/IIA/2
In some people, a gene mutation results in the formation of a defective digestive enzyme that may cause a
gastrointestinal disease.
One way to treat this disease is by enzyme replacement, i.e. introducing an external source of the digestive
enzyme into the patients’ gastrointestinal tract. The enzyme used can be produced by recombinant DNA
technology.
(c)(ii) Gene therapy can be an alternative method for the treatment of this genetic disease. State two
fundamental differences between the principles of gene therapy and enzyme replacement.
(2 marks)
(d) What potential hazards should scientists consider in the use of gene therapy in treating genetic
diseases? (3 marks)
Cloning
1. 2007/IC/12
Read the following passage and then answer the questions that follow it.
Plant and animal cloning
Plant and animal cloning is the production of genetically identical copies of 1
an individual. In agriculture and horticulture, the cloning of plants by artificial
propagation has a long history. With the advancement of technology,
traditional methods like cutting and grafting are gradually replaced by modern
methods involving tissue culture in the commercial production of horticultural 5
plants.
The cloning of plants by tissue culture is also known as micropropagation.
To achieve this, pieces of meristematic or non-meristematic tissues are taken
from the parent plant and grown under sterile conditions in special culture
media containing hormones. The cultured tissues will eventually develop into 10
plantlets which are then grown into whole plants in greenhouses.
In addition, tissue culture also makes it possible to generate transgenic
plants from a few cells, which have been genetically modified by recombinant
DNA technology. The production of transgenic plants by cloning has great
implications in agriculture and plant biotechnology. For instance, transgenic 15
cotton plants expressing a toxic bacterial protein are insect resistant. This is
because the expressed protein in plant tissues would specifically kill the insects
that feed on them.
Similarly, animal cloning has great potential applications in animal
biotechnology such as the production of superior animal stocks. Animal cloning 20
can also be used to propagate transgenic farm animals that can produce in their
milk pharmaceutical proteins such as human growth hormones. Furthermore,
cloned human embryos are a potential source of stem cells for generating tissues
or organs for transplants. However, advancement in this area is restricted at
present due to ethical concerns. Likewise, there are controversies and debates 25
about the development and the applications of animal and plant cloning,
including the production of transgenic organisms, despite their great potential.
(a) State two advantages of using meristematic tissues in tissue culture. (lines 6-9) (2 marks)
(b) Name one plant hormone that is usually used in the culture medium and state its importance.
(lines 6-9) (2 marks)
(e) Suggest three reasons why micropropagation has gradually replaced the traditional methods of cutting
and grafting in the commercial propagation of horticultural plants.
(3 marks)
(f) Compared with traditional breeding programs, discuss the advantages of cloning in producing
superior animals. (lines 18-19) (3 marks)
(g) Discuss the ethical concerns of cloning human embryos for medical research. (lines 22-24)
(4 marks)
 HKALE BIOLOGY | 29 Applied Genetics | P.4

Others
1. 1996/II/11
With regard to advances in molecular biology, describe and discuss the impact of the technological
manipulation of human genes on mankind. What are the ethical implications of this technology?
(20 marks)