CAT REVIEW & REVISION

TIPS

George Ichoho
 First Some Remarks
• Overall class performance was encouraging
• Many displayed a great understanding of key concepts especially in
the essay segment
• If you feel you scored lower than expected, do not be discouraged as
even those who didn’t score high showed a decent level of
understanding and can revise between now and the exam date to
score very high in the exam. The potential is evident from this CAT
 Revision Tips for MCQs
• Answer all questions: There is no negative marking
• Attempt to think of the answer before reviewing the options. This will
assist you in eliminating the incorrect options easier
• Go with your instincts: Several changed answers from the correct
ones noticed
• Details matter: Many picked answers that were not far off from the
correct answer e.g. question 2 of the MCQ which will be shown in a
few slides. It shows you did study the topic but these details matter
 Revision Tips for Essay Questions
• Always start with the definition
• For example in the question “Describe the Base Excision Repair (BER)
Process”, do not immediately go into the description. Break it down
into: definition, base excision repair, conclusion/summary if needed.
• Stick to the question asked. Non-pertinent information will not result
• Number of marks on the question dictates the level of detail
expected.
• Come up with your own Essay Questions and write your own answers
as practice to see if you fully understand a topic
 MCQs
1. Which nitrogenous base is not found in RNA?
A.Thymine
B. Adenine
C. Cytosine
D. Guanine
E. Uracil
 MCQs
2. DNA Replication is
A. Conservative and Unidirectional
B. Conservative and Bidirectional
C. Dispersive and Unidirectional
D. Semiconservative and Unidirectional
E. Semiconservative and Bidirectional
Three Possible Principles
Bidirectional Replication
 MCQs
3. Which structure is commonly referred to as beads on a string?
A. Chromosome
B. Plasmid
C. Ribosome
D.Chromatin
E. Lysosome
Chromatin: Beads on a String
 MCQs
4. What is the function of DNA Ligase during the DNA Replication process?
A. Prevents elongating DNA Polymerases from dissociating from the DNA parent
strand
B. Re-anneals the semi-conservative strands and joins Okazaki Fragments of the
lagging strand
C. separates the two strands of DNA at the Replication Fork behind the
Topoisomerase
D. Relaxes the DNA from its supercoiled nature
E. Provides a starting point of RNA (or DNA) for DNA Polymerase to begin synthesis
of the new DNA strand
 Replication Summary
Enzyme Function in DNA Replication
DNA Helicase AKA Helix destabilising enzyme. Helicase separates the two strands of DNA at
the Replication Fork behind the topoisomerase

DNA Polymerase Catalyses the addition of nucleotide substrates to DNA in the 5’ to 3’ direction
during DNA replication. Also performs proof-reading and error-correction.
There exist several types of DNA Polymerase, each of which perform different
functions in different types of cells

DNA Clamp A protein which prevents elongating DNA Polymerases from dissociating from
the DNA parent strand
ssDNA-Binding Protein Bind to ssDNA and prevents the DNA double helix from re-annealing after DNA
helicase unwinds it, thus maintaining the strand separation, and facilitate the
synthesis of the new strand
Topoisomerase Relaxes the DNA from its supercoiled nature
DNA Gyrase Relieves strain of unwinding DNA Helicase; this is a specific type of
topoisomerase
DNA Ligase Re-anneals the semi-conservative strands and joins Okazaki Fragments of the
lagging strand
Primase Provides a starting point of RNA (or DNA) for DNA Polymerase to begin
synthesis of the new DNA strand
Telomerase Lengthens telomeric DNA by adding repetitive nucleotide sequences to the
end of eukaryotic chromosomes. This allows germ cells and stem cells to avoid
the Hayflick limit on cell divisions
 MCQs
5. Which statement is incorrect?
A. RNA primers are longer in eukaryotes than in prokaryotes
B. DNA synthesis is primed by RNA
C. DNA replication is semi-conservative with a very low error rate
D.Okazaki fragments are longer in prokaryotes than in eukaryotes
E. Replication occurs in one direction
Bidirectional Replication
 MCQs
6. Alternative splicing
A. occurs in the cytoplasm
B. involves the removal of exons from pre-mRNA
C. allows the production of different proteins from a single gene
D.occurs prior to transcription
E. is one cause of gene silencing
 Alternative Splicing in Eukaryotes

➢ Although there are “only” 20-25,000 protein-coding genes in humans the

number of different proteins is far higher
➢ Pre-mRNAs can be spliced in different ways to produce different mRNAs and
therefore different proteins
 MCQs
7. Chromatin-remodelling complexes provide access to DNA for other
proteins by
A. recruiting other enzymes
B. modifying the N-terminal tails of core histones
C. denaturing the DNA by interfering with hydrogen-bonding between
base pairs
D.using the energy of ATP hydrolysis to slide the nucleosomes
E. proteolysis of core histones
 Nucleosome Structure is Dynamic

➢ In order to rapidly gain access to DNA, nucleosomes can be slid along

the DNA by: CHROMATIN-REMODELLING COMPLEXES
➢ Cycles of ATP hydrolysis push DNA along histone core, exposing the
DNA
 MCQs
8. What is the role of the spliceosome?
A. To fuse peptide fragments to form complete proteins
B. To remove introns from pre-RNA
C. To remove exons from pre-mRNA
D.To export mRNA from the nucleus
E. To direct RNA polymerase to promoters
 Splicing is Carried Out by The SPLICEOSOME

http://www.youtube.com/watch?v=FVuAwBGw_pQ&feature=related
 MCQs
9. In bacteria the role of sigma (σ) factors are to:
A.direct RNA polymerase to promoter elements
B. catalyse transcription termination
C. cause attenuation
D.stimulate mRNA degradation
E. direct ribosomes to start codons
Bacterial RNA Pol Requires SIGMA (s) to recognise Promoters

➢ Bacterial “core” RNA Pol (5 protein subunits) is sufficient for

transcription elongation but it cannot recognise promoters
➢ A sixth subunit called SIGMA (s) allows RNA Pol to recognise
promoters
➢ Sigma + core RNA Pol = HOLOENZYME
➢ The 2 key roles of sigma are to:
1. To direct RNAP to promoter elements, allowing sequence specific
binding
2. Stimulate the melting of DNA (open complex formation)
➢ Sigma is not required for elongation and DISSOCIATES soon after RNA
Pol leaves the promoter
 MCQs
10. Which of the following processes does not occur in bacteria?
A. Translation
B. Transcription
C. Splicing
D.Replication

E. Supercoiling
 MCQs
11. The polymerization of the gel used in PAGE occurs between
polyacrylaminde and:
A. N,N-acrylamide
B. Bisarylamide
C. Ammonium persulfate
D.N,N-metylenebisacrylamide
E. Tetramethylethylenediamine
 MCQs
12. If DNA is digested by endonucleases in four sites giving rise to
fragment of which two are equal in length, how many bands would be
seen after electrophoresis?
A. 3
B. 4
C. 5
D.6
E. 2
 MCQs
13. The fluorescent dye such as Ethidium is used for visualization of
DNA. How does Ethidium bind to DNA?
A. Stacked between histone molecule
B. Binds to the nucleoside base
C. Intercalated between the stacked bases
D.Binds to the phosphodiester backbone
E. Ethidium bromide possesses UV absorbance and can absorb energy
from nucleotides excited by absorbance of 260 nm radiation.
 MCQs
14. Which of the following will migrate faster? The condition is the
molecular weight of the following is equal:
A.Supercoiled circular DNA
B. Nicked circular DNA
C. Single stranded DNA
D.Double stranded DNA
E. Denatured protein
 MCQs
15. During DNA cloning, which of the following is not a crucial
requirement:
A. DNA inserts
B. PCR amplification
C. Vector
D.Protein expression
E. Restriction endonuclease
 MCQs
16. Transformation does not involve the following:
A. Cutting
B. Recombination
C. Heat shock
D.Propagation
E. Expression
 MCQs
17. Which of the following is not an essential feature for being a perfect
vector:
A. Origin of replication
B. Selectable marker
C. Multiple cloning site
D.Restriction site
E. Virulent gene
 MCQs
18. What is the major difference between cloning vectors and primary
vectors?
A. Selectable marker
B. DNA inserts
C. Presence of promotor
D.Presence of two Ori
E. Presence of a cloning site
 MCQs
19. Plasmid clones can be screened by the following:
A.By selectable markers
B. By bacterial resistance gene
C. For restriction site
D.By ARS sequence
E. PAGE
 MCQs
20. How many restriction sites are contained by a typical cloning
plasmid?
A. 1
B. 2
C. 3
D.None
E. More than 1
 Essay Questions
1. Describe the Base Excision Repair (BER) Process (10 Marks)
• Base excision repair (BER) is a DNA Repair Pathway that corrects
DNA damage from oxidation, deamination and alkylation.
• Such base lesions cause little distortion to the DNA helix structure.
• BER is initiated by a DNA glycosylase that recognizes and removes
the damaged base, leaving an abasic site that is further processed
by short-patch repair or long-patch repair that largely uses
different proteins to complete BER
 Essay Questions
• This process involves a whole host of enzymes called DNA
glycosylases, each of which can recognize a specific type of altered
base in DNA and catalyze its hydrolytic removal
• There are at least six types of DNA glycosylases, including those that
remove deaminated Cs, deaminated As, different types of alkylated or
oxidized bases, bases with opened rings, and bases in which a
carbon–carbon double bond has been accidentally converted to a
carbon–carbon single bond
BER
 Essay Questions
2. List 5 applications of molecular biology and recombinant DNA
technology in medicine (10 Marks)
Definition: Recombinant DNA (rDNA) is a technology that utilizes
enzymes to cut and paste together DNA sequences of interest. The
recombinant DNA sequences can then be placed into vehicles known as
vectors that ferry the DNA into a suitable host cell where it can be
copied or expressed.

Lots of examples: Agriculture, insulin production, genetic engineering,

tackling environmental issues etc.
Role of Recombinant DNA Technology to Improve Life Khan et al. 2016
 Essay Questions
• What is PCR? Discuss the principal and applications of PCR (10 Marks)

• Definition: Polymerase chain reaction (PCR) is a common laboratory technique used to make many
copies (millions or billions!) of a particular region of DNA. This DNA region can be anything the experimenter
is interested in. For example, it might be a gene whose function a researcher wants to understand, or a
genetic marker used by forensic scientists to match crime scene DNA with suspects.
 Essay Questions
1. Denaturation (96 °C): Heat the reaction strongly to separate, or denature, the DNA strands. This provides
single-stranded template for the next step.

2. Annealing (55-65 °C): Cool the reaction so the primers can bind to their complementary sequences on the
single-stranded template DNA.

3. Extension (72 °C): Raise the reaction temperatures so Taq polymerase extends the primers, synthesizing
new strands of DNA.

• This cycle repeats 25-35 times in a typical PCR reaction, which generally takes 2-4 hours, depending on the
length of the DNA region being copied. If the reaction is efficient (works well), the target region can go
from just one or a few copies to billions.

• That’s because it’s not just the original DNA that’s used as a template each time. Instead, the new DNA
that’s made in one round can serve as a template in the next round of DNA synthesis. There are many
copies of the primers and many molecules of Taq polymerase floating around in the reaction, so the
number of DNA molecules can roughly double in each round of cycling. NB: Know the difference between
PCR & Sequencing. They are not the same
 Exam: What to expect
• More of the same format (MCQs & Essay Questions)

• Every lecturer gets to submit questions for both MCQs and Essay

• For my lectures: email me any queries you may have please email
them and I will get back to you ASAP
 Good Websites to Use During Revision
• Khan Academy: Covers a wide range of Molecular Biology topics with
good summaries and visual diagrams