ANALYSIS PAST YEAR QUESTION

i.

DNA RECOMBINANT TECHNOLOGY

QUESTION
1. Describe what is meant by triparental
mating? (15m)
2. In a triparental mating experiment, the
helper strain, host cell and the vector
used are resistant to ampicillin,
tetracycline and chloramphenicol,
respectively. Suggest 1 essential
antibiotic named above to select the
transconjugants. (5m)
3. How often would you expect to find a
ATCG restriction site in agenomic base
composition of 50% G+C? Show your
calculation. (1m)
How often would you expect to find a
GGCC restriction site in agenomic base
composition of 55% G+C? Show your
calculation. (5m)
How often would you expect to find a
GAGTC restriction site in agenomic base
composition of 45% G+C? Show your
calculation. (5M)
-State 3 assumptions that you have
made in order to solve the 3 questions
above. (9m)
4. -Explain the role of electrophoresis in
gene cloning. (10m)
-Explain how to isolate and purify a
sample of DNA from bacteria. (10m)
5. Discuss the facrors for consideration in
DNA Southern hybridisation. (10m)
-Explain the possible reasons why a DNA
Southern hybridisation fails? (10m)
6. Name the essential components for
polemerase chain reaction (PCR). (10m)
-Explain briefly why it is important to
perform negative control in PCR. Explain
the possible reasons why PCR fails (10m)
7. It is a common practice to grow
transformant using selection medium
supplement with appropriate antibiotic.

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You realised that the edu gene confers

ampicillin resistance to E. coli, and the
host cell itself is naturally resistant to
kanamycin, explain why neither
ampicillin and kanamycin-enriched
selection medium is not appropriate for
this purpose?
8. -Descibe the principle of NextGeneration Sequencing in detail. In
your opinion, what is the major
drawback of this technology? (10m)
-compare and contrast between NextGeneration Sequencing and Sanger
sequencing methods. (5m)
9. Explain critically the differences
between DNA replication in the cell and
Polymerase Chain Reaction. (10m)
-Give 2 reasons why this pairs of primers
isnt suitable for PCR.(5m)
10.
Explain a method for mutagenesis
in bacteria. (15m)
11.
List down the methods of
introducing sticky ends to blunt ends
molecues (3m)
12.
Define the following enzymes and
explain how are they useful in molecular
biology. (9m)
13.
Explain a method how you can
ensure ckg is constitutively expressed?
14.
Explain 1 main reason why not
many mutants will be obtained in
bacteria mutagenesis. (2m)
15.
You are studying a prokaryote
gene named C. Explain in detail, how
you would:
-isolate the gene C from bacterium (5m)
-ensure that the gene C has been cloned
into the plasmid vector (5m)
-determine that the gene C is actively
expressing (5m)
16.
Explain 1 technique of site directed
mutagenesis. (13m)
17.
Construct a restriction map of
linear fragment of DNA, using the

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following data. (5m)

18.
The genetic and physical map of
Pbr322 gives an indication why this
plasmid was such a popular cloning
vector. Elaborate n detail the 3 useful
properties of Pbr322. (9m)
19.
Discuss why sticky ends increase
the efficiency of ligation. (6m)
20.
Explain how Klenow fragment was
obtained and how it differs from DNA
polymerase I? (7m)
21.
Describe all the major types pf
plasmid classes according to their main
characteristics coded by the plasmid
genes. (5m)
22.
Explain in detail the steps involved
in Southern Blotting and Hybridization.
(8m)
23.
Explain the methods of introducing
sticky ends to blunt ended molecules
(12m)
24.
Mrna itself cannot be ligated into a
cloning vector. Explain how to overcome
this problem? (4m)
25.
Define isoschizomer and
neoschizomer and give examples of
each type of enzyme. (4m)
26.
There are a variety of strategies
available for the delivery of genes into
eukaryotic cells. Explain 4 methods of
transformation that are commonly used.
(8m)
27.
Explain why insertional
inactivation does not always involve
antibiotic resistance. (6m)
28.
Construct a restriction map of the
10kb DNA. (6m)
29.
Explain the 4 mehods of labelling
probes. (8m)
30.
The procedure for total DNA
preparation from bacteria culture can be
divided into 4 stages. Describe all the
stages. (4m)

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31.
Describe in detail the 5 basic steps
in a gene cloning experiment. (5m)
32.
Discuss the 3 parameters affecting
hybridization strategy in Southern blot.

Jan 2013, Jun 2013

Mei 2010, Nov
2010, Jan 2013,
Jun 2013
Jun 2013

33.
Construct a restriction map of a
circular DNA plasmid.
34.
Draw a genetic map that describes Jun 2013
the order of the three genes and the
distances between them. (12 m)
35.
Does chiasma interference occur in Jun 2013
this crossing-over? If yes, state whether
it is a positive or a negative type. (3m)
36.
When two populations frequently
Jun 2013
intermix due to migration, what are the
long-term consequences with regard to
allele frequencies and genetic variation?
Give appropriate to substantiate your
answer. (15m)

ii.

IMMUNOLOGY

QUESTION
1. Describe the genetic basics for the large
number of different antibody specificities
achieved. (10m)
2. Describe the genetic basics for the
different isotypes of the same specificity
produced. (5m)
3. Describe the genetic basics for allelic
exclusion to ensure expression of single
antigenic specificity. (5m)
4. Discuss the advantages for the existence

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of both the innate and adaptive immune

systems in enhancing protection against
infection. (20m)
5. What are cytokines? (2m)
6. Describe the biological activities of
cytokines with examples to show their
major roles in the immune system.
(18m)
7. What is MHC restriction? (2m)
8. Describe the function of the thymus in
the development of a mature T cell.
(18m)
9. Describe the antigen binding site of MHC
class I. (4m)
10.
What are the functions of the
invariant chain and CLIP in the MHC
class II exogeneous pathway? (6m)
11.
What are the maximum number of
class I and class II proteins expressed on
a liver cell? On a macrophage? (4m)
12.
Why might certain MHC alleles be
associated with certain diseases?
13.
The two major subpopulations of T
lymphocytes are the CD4T helper cells
and CD8T cytotoxic cells. Describe how
these two subpopulations of T
lymphocytes regulate the immune
system? (5m)
14.
The T cell is said to be class I
restricted. What does this mean? (5m)
15.
Positive selection involves
selection for thymocytes bearing
receptors capable of binding self-MHC
molecules. Why do such large numbers
of thymocytes fail positive selection?
(5m)
16.
Negative selection involves
elimination of thymocytes bearing high
affinity receptors for self-MHC molecules
alones of self-antigen presented by selfMHC. Which thymus cells determine self
for negative selection? (5m)
17.
Describe internalization,
processing and presentation of an

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endogenous antigen by a B cell. (20m)

18.
How do cytokines transmit their
information to cells during an immune
response? (5m)
19.
When IL-2 is screted by one T cell
in a peripheral lymphoid organ, do all T
cells in the vicinity proliferate in
response to the IL-2 or only some of
them? (5m)
20.
Therapies based on cytokines and
cytokine receptor have entered clinical
practice, one important cytokine being
interferon. Discuss the current success
of interferon in treatment of diseases.
(10m)
21.
Describe the effector mechanisms
that are classified as adaptive immunity.
(20m)
22.
There are two phases in the
development of B cells; an antigenindependent phase involving maturation
and an antigen-dependent phase
involving activation and differentiation.
Describe the events in B cell
development that are antigen
independent. (20m)
23.
How does one acquire innate
immunity? (4m)
24.
Give short notes on how these
innate cells function to provide an
immune response:
-macrophages
-neutrophils
-dendritic cells
-natural killer cells
(8m)
25.
Redness and swelling are typical of
inflammatory responses. What are
mechanism s involved in these
responses? (4m)
26.
What are the benefits of redness
and swelling? (4m)
27.
What is the unique feature about
polymorphism of class I and ll MHC

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genes? What is the advantage of this

unique feature? (4m)
28.
Predict whether TH cell
proliferation or CTL-mediated cytolysis or
n responses of target cells will occur with
the following mixtures of cells. The
CD4TH cells are from lysozyme-primed
mice, and the CD8+CTLs ARE from
influenza-infected mice. Briefly explain
your answer. (12m)
29.
A Tc cell clone recognises a
particular measles virus peptide-binding
cleft identical to the one in H-2Db. A
second MHC molecule has a peptidebinding cleft identical to the ine in H-2Db
except differs at several other amino
acids in the X1x2 domain. Predict
whether the second MHC molecule
coukd present this measles virus peptide
to the Tc cell clone. Briefly explain your
answer. (4m)
30.
Explain why serum IgM cannot
activate complement by itself.(2m)
31.
Describe 3 effector functions of
complement that help the body combat
microbial infection? (15m)
32.
Why are there so many
complement regulators (3m)
33.
In the process of genetic
rearrangement, why is allelic exclusion
important (3m)
34.
Why do we continuously produce
lymphocytes? (2m)
35.
The yo T cell receptor differs from
the xB in both structural and functional
parameters. Describe how they are
similar to one another and different from
the B cell antigen receptors. (3m)
36.
Which effector mechanisms are
classified as adaptive immunity? (12m)
37.
List the primary lymphoid organs
and briefly describe their functions in the
immune system. (6m)
38.
What is immune memory? Why is

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it important? (in ensuring success of the

immune system to protect a person
against infection from a virulent
pathogen)(4m)
39.
How does clonal selection explain
antigen specitivity, discrimination
between self and non-self and memory
characteristics of adaptive immunity?
(10m)
40.
Discuss 3 mechanisms in which
variation is introduced into T and B-cell
receptor during T and B-cell
development? (15m)
41.
T- and B-cells recombine their DNA
to create diversity rather than using
alternative splicing. Why do you think
this is so? (5m)
42.
Draw a secreted form of IgA. (6m)
43.
Name 4 antibody classes. (4m)
44.
What are the advantages of
changing an antibody class? (4m)
45.
What are the 3 effector functions
of antibody? (6m)
46.
What are the 4 features of the
adaptive immune system that allow it to
effectively fight a multitude of possible
infections and why is each necessary?
(8m)
47.
How do the jawless fishes and
invertebrates survive without an
adaptive immune system?(2m)
48.
What is the difference between an
antigen and an immunogen? (4m)
49.
Name 3 factors that affect the
immunogenicity of a protein? (6m)
50.
When an organ is transplanted, the
patient must take immunosuppressant
drugs for the rest of hid life. Why is that?
(5m)
51.
The exception to this is bone
marrow transplant. What makes bone
marrow different from, say, a kidney that
the patient does not require
immunosuppressant to live? (5m)

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52.
Name 1 complication of bone
marrow transplant that is generally not a
concern in other transplant, and how it is
avoided. (5m)
53.
Explain why, after a bone marrow
transplant, the patient must often get an
entire complement of vaccinations. (5m)
54.
Name 5 differences between
innate and adaptive immunity (10m)
55.
Nam3 5 major differences between
MHC l and MHC ll? (10m)
56.
What are the four main functions
of the innate immune system? (4m)
57.
Discuss in detail the 4 types of
defensives barriers of the innate immune
system. (12m)
58.
Most of the
antigen/pathogen/organisms
encountered by a healthy individual are
readily cleared within a few days by
defense mechanisms of the innate
immune system Why do you think this
is possible? (4m)
59.
In order to protect the body
against a diverse number of pathogens a
large number of antibodies with different
specificities, must be generated. How is
this achieved? (10m)
60.
Different isotypes of the same
specificity are being produced in
response to an infection, Why do you
think generating the same specificity in
an immune response is important?
Describe briefly the process that takes
place to ensure this. (4m)
61.
Complement activation can occur
via the classical, alternative, or lectin
pathways. How do the three pathways
differ in the components that can initiate
their activation? (3m)
62.
Which portion of the overall
activation sequence differs in the 3
pathways? Which portion is similar?(4m)
63.
How do the biological

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cinsequences of complement acticatiob

via these pathways differ> (6m)
64.
Would you expect a C1 or a C3
complement deficiency to be more
clinically serious? Describe briefly the
implication on the health of a person
with deficiency in C1 or C3. (7m)
65.
Explain briefly:
-Endogenous antigen
-Exogenous antigen
-Antigen processing
-Self-MHC restriction
66.
One of the characteristics features
of the MHC is a large number of different
alleles at each locus. Where are the
most polymorphic amino acis residues
locates in the MHC molecules? Describe
briefly on the importance of this
location? (4m)
67.
Why do you think it is important
for MHC molecules to be involved in the
selection of T cells in the thymus? (4m)
68.
What is the difference in immune
responses towards an extracellular
bacteria as compare to an intracellular
virus? (4m)

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