PART I (Dr.

Lam's part, total 60 marks)

(A) Answer any THREE questions below. Each carries 8 marks.

(1) What is the purpose of using a dut- ung- double mutant in primer extension experiments?
(2) In yeast functional complementation experiments, what are the major features of the yeast vector
involved?
(3) HAT medium contains hypoxanthine, aminopterin, and thymidine. Explain how the HAT
medium can help the gene transfer into mammalian cells.
(4) Mammal cloning is one major breakthrough in biotechnology recently. Briefly explain the basic
principles.
(5) Explain the basic principles of a binary Ti plasmid system in plant transformation.
(6) Many plant genes related to the developmental processes were cloned by the help of T-DNA
transformation. Explain.

(B) Answer ALL questions.

(1) The activation of vir genes in a Ti-plasmid required a two-component regulatory system
composed of VirA and VirG proteins. After VirA receives the plant wound signals, it will be
autophosphorylated. VirG, a transcription factor regulating other vir genes, is subsequently
phosphorylated by VirA via phosphate transfer.
(a) Design an experiment to show that there is a protein-protein interaction between VirA and VirG.
(8 marks)
(b) A specific histidine residue on the VirA protein is thought to be the site of phosphorylation.
Design an experiment to test this hypothesis. (4 marks)

(2) Glutamate receptors are a group of neuroreceptors in animals which use glutamate as the
neurotransmitter. When glutamate molecules bind to the glutamate receptors, a transient
electrical currents will be generated across the plasma membrane.
(a) Suppose you are the first scientist who attempts to clone the gene for a glutamate receptor. What
will be your experimental strategies. (6 marks)
(b) After you obtain the gene, design one experiment to test the in vivo physiological functions of the
gene inside an animal. (6 marks)

(3) The mRNA levels of a plant gene (Gene X) is much higher in the dark then in the light. You
would like to explore if there is an induction by the dark or a repression by the light. To test the
two possible models and to locate the regulatory site(s), you have cloned the promoter of Gene X
(500 bps), performed nested deletion, and fused all constructs to the reporter gene GUS. All GUS
constructs were then transformed into plants and the GUS activities were measured.

Promoter region GUS unit in Light GUS unit in Dark

-500 to +1 10 1000
-500 to -300 1 100
-500 to -100 10 1000
-400 to +1 10 1000
-300 to +1 1000 1000
-200 to +1 100 100

(a) Is Gene X regulated by light repression or dark induction? Where is the location of the
PART II (Prof. Sun's part, total 40 marks)

Please write answers for Part II in the answer booklet

Answer all the following questions

1. The Human Genome Project is an international, long term, and very expensive undertaking.
Why are we doing it? Give all the reasons that you can think of. (5 marks)

2. The scientists involved in the Human Genome Project had developed a research plan to
accomplish the project. Could you outline the key components of this plan? (5 marks)

3. As of today, in comparison to animal biotechnology, plant biotechnology has made notably

more progress in developing and commercializing biotechnological products. Suggest
possible reasons for this happening. (5 marks)

4. The first human clinical trial in gene therapy occurred in 1992 in the US. Amy Harper with
combined immunodeficiency disease was treated with the adenosine deaminase gene. Three
components are essential for such therapy. What are these components and why are they
essential? (5 marks)

5. There are many limitations in conventional vaccine production. Application of

recombinant DNA technology would help eliminate some of these limitations.
Explain this new technological capability in this regard with examples. (7 marks)

6. It has been demonstrated that transgenic plants can be used as bioreactors to produce useful
medical, food, and industrial products.

a) What is the molecular basis involved here? (5 marks)

b) What are the advantages of using plants as bioreactors instead of using bacterial,
yeast, or animal cells and animals? (5 marks)
c) Any disadvantages you could think of in using plants? (3 marks)

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