Student ID

University of Bedfordshire

Unit Code: BHS001-6

Unit Title Cell and Molecular Biology

Academic Year 2020-21

Period of Study SEM 1 Paper 2

Date of Exam: 13 April 2021

Start Time: 14:00

Time Allowed: 3 hours

Unit Co-Ordinator: Dr Prashanth Bajpe

Instructions for Candidates

Do not open this paper until instructed to do so by the Senior invigilator
Equipment allowed: Calculator

Materials allowed: None

Materials supplied None

Additional Instructions: Type of Exam:

This exams is an open /a closed book examination
 Please answer 3 questions in total.
There are two sections
 SECTION A: Answer TWO questions in this section (all questions in this section
are of equal weighting)
 SECTION B: Answer the compulsory question in this section
 Section A is worth 60% of the total mark (30% for each of the two questions)

 Section B is worth 40% of the total mark.

BHS001-6 /20-21/ SEM1/ Paper 2 Page 1 of 4

 SECTION A – Answer TWO questions in this section

This section is worth 60% of the total mark (30% for each of the two questions)

Question 1
a. Nowadays, diabetic patients can purchase human insulin from a pharmacy.
Critically discuss the technology that allows its production and the advantage of the
methodology over previous strategies (50%).
b. Using appropriate examples, elaborate the current and potential applications of
recombinant protein expression in animals or plants (50%).

Question 2
The field of functional genomics has been revolutionised by the discovery of RNA
interference.
a. Critically discuss this statement and provide example applications of this technology
(50%)
b. Compare RNA interference to knockout animal models in understanding the
functions of genes (50%)

Question 3

a. Describe the different components of the CRISPR-CAS system. Discuss why

CRISPR-CAS system is a revolutionary gene editing technology (40%)
b. Outline the role of two different DNA repair pathways in the CRISPR-CAS gene
editing system (35%).
c. In order to make knock outs of TP53 gene, CRISPR-CAS technology was used to
make changes to the sequence of TP53 DNA. A single guide RNA (sgRNA)
targeting TP53 gene and CAS9 protein was expressed in a cell line. Following this,
it was confirmed that TP53 gene sequence was edited (just one base removed by
CRISPR-Cas) leading to a frame shift mutation and as a consequence inclusion of
a premature stop codon (UAA). To confirm whether this edit can knockout/disrupt
TP53 gene, RT-PCR was performed to check the expression of TP53 mRNA.
Surprisingly, there was no difference in the expression of TP53 mRNA between
non-edited control cells and edited cells (check the figure below). Identify the error
that might have occurred in choosing this assay. Elaborate your answer by
describing the type of technique that needs to be used in this experiment in order to
confirm the knockout. (25%)
 TP53 mRNA expression in non-edited control cells and edited cells.

Question 4

a. Describe in detail eukaryotic DNA replication process in a step-by-step manner. (40%)

b. Critically evaluate the core factors involved in the formation of transcription preinitiation
complex. Include in your answer the recruitment of RNA polymerase II onto specific genes.
(40%)
c. Below is an RNA sequence with two introns (Intron 1 and Intron 2) and two exons (Exon
1 and Exon 2). Exonic sequences are coloured blue and intronic sequences are coloured
red. Provide the RNA sequence after splicing. (20%)

RNA sequence with Exon(blue) and Intron(red)

SECTION B (40% of total mark) – Answer the compulsory question in this section.

Compulsory Question
a. Critically discuss the principles of nucleic acid hybridisation and include in your
answer an example of molecular biology technique which is based on nucleic acid
hybridisation for detection of a mutation in a DNA sequence.(40%)
b. A PCR was performed by using the components shown in table 1 and the cycling
conditions shown in table 2. Recommend a technique that would allow you to check
if the reaction was successful. However, in case it was unsuccessful, suggest a few
optimisation strategies. (40%)

Table 1 components of PCR reaction mix

Component Concentration Volume
10x PCR buffer 5µl
dNTP mix 10mM 1 µl
Forward primer 10 µM 1 µl
Reverse primer 10 µM 1 µl
nH2O 39 µl
Template DNA 50ng 1 µl
Taq polymerase units 1 µl

Table 2 PCR cycling conditions

Temperature Time Number of
cycles
94oC 2 minutes 1
94oC 30 seconds
60oC 30 seconds 30
72oC 30 seconds
72oC 10 minutes 1
4 oC ∞ Hold

c. Explain how PCR can produce amplicons of different sizes by using the same pair
of primers. (20%)