2.

7 DNA REPLICATION,
TRANSCRIPTION AND TRANSLATION
Essential Idea
Genetic information in DNA can be accurately copied and
can be translated to make the proteins needed by the cell
UNDERSTANDINGS
UNDERSTANDINGS
INTRODUCTION
• During the S-
phase, prior to cell
division, DNA is
replicated to make
identical copies, so
that each daughter
cell is a perfect copy
of the parent cell
PHASE LOCATION IN ACTIVITIES
CELL
G1 (Gap 1) Cytoplasm • The cell grows and functions normally
undergoing everyday processes
• Rapid protein synthesis takes place
allowing the cell to grow in size
• Cell makes RNA, enzymes and other
proteins needed for growth and for DNA
synthesis (the next phase)
• Additional organelles are made

S (synthesis) Nucleus • The amount of DNA doubles as DNA

replication takes place, going from 2n
to 4n in preparation for nuclear division
• The genetic material is duplicated but
no chromosomes are formed yet
INTRODUCTION
• DNA replication is a
high precision
exercise that involves
a set of enzymes
working in a
coordinated way to
produce perfect
daughter molecules
DNA REPLICATION
Understanding
The replication of DNA is semi-conservative and depends on
complementary base pairing
DNA REPLICATION
•It is the precise
and accurate
copying or
duplication of DNA
or genetic material
DNA REPLICATION
• The Watson-Crick
model of the DNA
structure allowed for
a logical explanation
of how DNA
produces exact
copies of itself
DNA REPLICATION
•The Watson-Crick
model led to the
semi conservative
replication
hypothesis for DNA
replication
DNA REPLICATION
• Therefore, three main
hypotheses were
proposed to explain the
mechanism of replication
namely:
• Conservative replication
• Semi-conservative
replication
• Dispersive replication
POSSIBLE MECHANISMS OF DNA REPLICATION
Mechanisms of DNA replication

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THE MESELSON AND STAHL
EXPERIMENTS
Nature of Science
Obtaining evidence for scientific theories—Meselson and Stahl
obtained evidence for the semi-conservative replication of DNA
Skill
Analysis of Meselson and Stahl’s results to obtain support
for the theory of semi-conservative replication of DNA
THE MESELSON AND STAHL EXPERIMENTS
• Meselson and Stahl
designed some
elegant experiments,
which demonstrated
beyond doubt that
DNA replication
progresses in a semi-
conservative way
THE MESELSON AND STAHL EXPERIMENTS

•They cultured
E.coli bacteria in
the presence of a
heavy nitrogen
isotope,  N
15
THE MESELSON AND STAHL EXPERIMENTS
• DNA contains nitrogen
in its nitrogenous
bases, so the
radioactive 15N would
end up in the DNA of
the bacteria, hence all
bacterial DNA had 15N
in its bases
THE MESELSON AND STAHL EXPERIMENTS
• They then transferred the
bacterial culture into a
fresh medium where the
nitrogen was replaced
by 14N, a lighter isotope,
and the bacteria were
allowed to grow for
several generations
THE MESELSON AND STAHL EXPERIMENTS
• DNA samples were then
extracted from
successive bacterial
generations and
subjected to
caesium chloride
equilibrium density
gradient centrifugation
THE MESELSON AND STAHL EXPERIMENTS
• This technique
allowed the DNA to
move to different
positions in the
centrifuge tube
based on its density
THE MESELSON AND STAHL EXPERIMENTS
• DNA containing one
or two strands with 15N
was heavier and showed
lower bands than those
containing two strands
with 14N (DNA with 15N in
both strands was
heaviest).
THE MESELSON AND STAHL EXPERIMENTS
• After one generation, i.e.
one division of
the bacteria,
the resulting DNA strand
consisted of a double
helix, where one strand
was made up of 15N, and
the other contained 14N
THE MESELSON AND STAHL EXPERIMENTS
• The band obtained
was in between
those for DNA with
both strands
containing 15N or
14
N only
THE MESELSON AND STAHL EXPERIMENTS
• This indirectly
demonstrated that DNA
replication had to proceed
in a semi-conservative way
and should
involve complementary
base pairing to ensure the
fidelity of the daughter
molecules
Meselson–Stahl DNA experiment

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SEMI-CONSERVATIVE
REPLICATION OF DNA
Understanding
The replication of DNA is semi-conservative and
depends on complementary base pairing
SEMI-CONSERVATIVE REPLICATION OF DNA
• DNA replication is a semi-
conservative process,
because when a new
double-stranded DNA
molecule is formed:
• one strand will be from the
original template molecule
• one strand will be newly
synthesised
SEMI-CONSERVATIVE REPLICATION OF DNA
• Thus DNA replication is
semi-conservative as
each daughter molecule
formed contains one
original strand from the
parent molecule and
one newly synthesised
strand
SEMI-CONSERVATIVE REPLICATION OF DNA
• This occurs because
each nitrogenous base
can only pair with its
complementary partner
• Adenine (A) pairs with
Thymine (T)
• Cytosine (C) pairs with
Guanine (G)
SEMI-CONSERVATIVE REPLICATION OF DNA
• Consequently, when DNA is
replicated:
• Each new strand formed will
be identical to the original
strand separated from the
template
• The two semi-conservative
molecules formed will have
an identical base sequence to
the original molecule
THE MECHANISM OF DNA
REPLICATION
Understandings
THE MECHANISM OF DNA REPLICATION
•The process of
DNA replication is
coordinated by
two key enzymes:
• helicase and
• DNA polymerase
THE MECHANISM OF DNA REPLICATION
•Helicase unwinds
the double helix
and separates the
two polynucleotide
strands
THE MECHANISM OF DNA REPLICATION
•It does this by
breaking the
hydrogen bonds
that exist between
complementary
base pairs
THE MECHANISM OF DNA REPLICATION
•The two separated
polynucleotide
strands will act as
templates for the
synthesis of new
complementary
strands
THE MECHANISM OF DNA REPLICATION
•DNA polymerase
synthesises new
strands from the
two parental
template strands
THE MECHANISM OF DNA REPLICATION
•Free deoxynucleoside
triphosphates
(nucleotides with 3
phosphate groups)
align opposite their
complementary base
partner
THE MECHANISM OF DNA REPLICATION
•DNA polymerase
cleaves the two
excess phosphates
and uses the energy
released to link the
nucleotide to the
new strand
NOTE
• As the two DNA strands of the double helix are anti-
parallel to each other, DNA polymerase proceeds in
opposite directions during replication
• On one strand it moves in the same direction as the
replication fork (immediately behind the helicase
enzyme), and moves in the reverse direction on the
other strand
• DNA replication always occurs in the 5' to 3' direction
DNA replication

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DNA replication

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SYNTHESIS OF PROTEINS
SYNTHESIS OF PROTEINS
•Enzymes are
proteins that
control reactions
involved in the
development of
organisms
SYNTHESIS OF PROTEINS
• A gene is a specific
sequence of
nucleotide bases on a
DNA molecule that
codes for a
polypeptide (protein)
or a functional mRNA
SYNTHESIS OF PROTEINS
•Therefore
genes control
the nature and
development of
all organisms
SYNTHESIS OF PROTEINS
• Protein synthesis is
the assembling of
free amino acid
units in the cell into
a functional
polypeptide chain or
protein
SYNTHESIS OF PROTEINS
•The process of
protein synthesis
involves two main
stages:
•Transcription
•Translation
TRANSCRIPTION
Understanding
Transcription is the synthesis of mRNA copied from
the DNA base sequences by RNA polymerase
TRANSCRIPTION
•It is the synthesis
of mRNA copied
from the DNA
base sequences
by RNA
polymerase
TRANSCRIPTION
•It is the process
of making mRNA
using one strand
of the section of
DNA as a
template
TRANSCRIPTION
• When a gene is
transcribed, the
DNA forms a
template for the
RNA polymerase to
synthesise (make) a
copy of mRNA
THE MECHANISM OF
TRANSCRIPTION
Understanding
Transcription is the synthesis of mRNA copied from
the DNA base sequences by RNA polymerase
THE MECHANISM OF TRANSCRIPTION
•RNA
polymerase
binds to a site
on the DNA at
the start of a
gene
THE MECHANISM OF TRANSCRIPTION
• RNA polymerase moves
along the gene,
separating DNA into
single strands and
pairing up RNA
nucleotides with
complementary bases on
one strand of the DNA
THE MECHANISM OF TRANSCRIPTION
•There is no
thymine in RNA,
so uracil pairs in a
complementary
fashion with
Adenine
THE MECHANISM OF TRANSCRIPTION
• RNA polymerase
removes the additional
phosphate groups from
free ribonucleoside
triphosphates and uses
the energy from this
cleavage to covalently
join the RNA nucleotides
to the growing sequence
THE MECHANISM OF TRANSCRIPTION
• Once the RNA
sequence has been
synthesised, RNA
polymerase detaches
from the DNA
molecule and the
double helix reforms
THE MECHANISM OF TRANSCRIPTION
•Transcription
stops at the end of
the gene and the
completed mRNA
molecule is
released
NOTE
•Transcription
proceeds in the 5’- 3’
direction
NOTE
•RNA polymerase is responsible
for both separating the DNA strands of
the double helix as well as for joining
the ribonucleotides together by
phosphodiester bonds to form an
mRNA strand
THE MECHANISM OF TRANSCRIPTION
•The transcribed
strand of the DNA
molecule is known
as the antisense
strand and is
complementary to
the mRNA molecule
THE MECHANISM OF TRANSCRIPTION
• The DNA strand that is
not transcribed is called
the sense strand
and has the same
sequence of bases as
the mRNA molecule,
except for thymine being
replaced by uracil
THE MECHANISM OF TRANSCRIPTION
• Once mRNA is
made, it leaves the
nucleus and
attaches to a
ribosome where it
forms the template
for protein synthesis
CODON
Understanding
The amino acid sequence of polypeptides is
determined by mRNA according to the genetic code
CODON
•It is a
sequence of
three bases
on an mRNA
molecule
CODON
•Each codon
codes for one
amino acid
within a
polypeptide
chain
CODON
•Each codon of
three bases on the
mRNA is translated
into one amino acid
in a polypeptide
chain
CODON
• The order of the
codons in an mRNA
sequence
determines the
order of amino
acids in a
polypeptide chain
THE GENETIC CODE
Understanding
The amino acid sequence of polypeptides is determined by
mRNA according to the genetic code
Codons of three bases on mRNA correspond to one amino
acid in a polypeptide
THE GENETIC CODE
• The genetic code is the
set of rules by which
information encoded
within mRNA sequences
is converted into amino
acid sequences
(polypeptides) by living
cells
THE GENETIC CODE
• It is the order of
codons that
determines
the specific
sequence of amino
acids in a
polypeptide chain
THE GENETIC CODE
• It is the sequence of
bases in mRNA for
putting together
amino acids in the
correct order to a
make a specific
protein
THE GENETIC CODE
• The code is a three-
letter, or triplet,
code
• Each sequence of
three bases stands
for one amino acid
THE GENETIC CODE
•Two amino
acids are
coded for by
only a single
triplet
THE GENETIC CODE
•The remaining
amino acids are
coded for by
between 2 to 6
triplets each
THE GENETIC CODE
• The code is a
degenerate code
which means that
most amino acids
are coded for by
more than one
triplet
THE GENETIC CODE
•The code is
read in the
5’ → 3’
direction
THE GENETIC CODE
•The start of a
sequence is
always the triplet
AUG which codes
for the amino acid,
methionine
THE GENETIC CODE
• Three codons, UAA,
UAG, and UGA, do
not code for any
amino acid and are
called STOP codons
which mark the end of
a polypeptide chain
THE GENETIC CODE
•The genetic code
is non-
overlapping, i.e.
each base in the
sequence is read
once
THE GENETIC CODE
• The code is
universal i.e. each
triplet codes for the
same amino acids
in all organisms
with a few
exceptions
HOW TO USE THE GENETIC CODE TO
DEDUCE WHICH CONDONS
CORRESPOND TO AMINO ACID
Skill
Use a table of the genetic code to deduce which
codon(s) corresponds to which amino acid
HOW TO USE THE GENETIC CODE TO DEDUCE WHICH
CONDONS CORRESPOND TO AN AMINO ACID

•Identify the
amino acid
from the
genetic code
HOW TO USE THE GENETIC CODE TO DEDUCE WHICH
CONDONS CORRESPOND TO AN AMINO ACID
• Look down the left
hand side of the table
to find the first base of
a codon, across the
top to find the second
base and down the
right hand side to find
the third base
TRIAL QUESTION
TRIAL QUESTION
HOW TO USE THE CONDONS TO
DEDUCE THE SEQUENCE OF AMINO
ACIDS
Skill
Use a table of mRNA codons and their corresponding amino
acids to deduce the sequence of amino acids coded by a short
mRNA strand of known base sequence
HOW TO USE THE CONDONS TO DEDUCE
THE SEQUENCE OF AMINO ACIDS
• Look down the left
hand side of the table
to find the first base of
a codon, across the
top to find the second
base and down the
right hand side to find
the third base
HOW TO USE THE CONDONS TO DEDUCE
THE SEQUENCE OF AMINO ACIDS
• Determine the
amino acid that
corresponds to the
base sequence i.e.
all three base from
the left to the right
TRIAL QUESTION
TRIAL QUESTION
HOW TO USE THE DNA SEQUENCE TO
DEDUCE THE SEQUENCE OF mRNA
Skill
Deducing the DNA base sequence for the mRNA strand
HOW TO USE THE DNA SEQUENCE TO DEDUCE THE
SEQUENCE OF mRNA

•A strand of mRNA
is produced by
transcribing the
anti-sense strand
of the DNA
molecule
HOW TO USE THE DNA SEQUENCE TO DEDUCE THE
SEQUENCE OF mRNA

•The anti-sense
strand therefore is
complementary to
the mRNA
TRIAL QUESTION
TRIAL QUESTION
UNIVERSALITY OF THE
GENETIC CODE
Application
Production of human insulin in bacteria as an example of
the universality of the genetic code allowing gene transfer
between species
UNIVERSALITY OF THE GENETIC CODE
• The universality of the
genetic code means
that every living
organism uses the
same code with a few
rare and minor
exceptions
UNIVERSALITY OF THE GENETIC CODE
• As the same
codons code for the
same amino acids
in all living things,
genetic information
is transferrable
between species
UNIVERSALITY OF THE GENETIC CODE
• The ability to transfer
genes between
species has been
utilised to produce
human insulin in
bacteria for mass
production
INDUSTRIAL INSULIN
PRODUCTION
Application
Production of human insulin in bacteria as an example of
the universality of the genetic code allowing gene transfer
between species
INDUSTRIAL INSULIN PRODUCTION
•mRNA for insulin is
extracted from
pancreatic β cells
as they are the only
cells to express the
insulin gene
INDUSTRIAL INSULIN PRODUCTION
•The mRNA is
then incubated
with the enzyme
reverse
transcriptase
INDUSTRIAL INSULIN PRODUCTION
• This enzyme
reverses
transcription, using
mRNA as a
template to make
single stranded
DNA
INDUSTRIAL INSULIN PRODUCTION
• These single-stranded
DNA molecules are then
converted to double-
stranded DNA
molecules using DNA
polymerase to assemble
nucleotides to make the
complementary strand
INDUSTRIAL INSULIN PRODUCTION
• The insulin genes are
now inserted into
plasmids to transform
the bacterium
Escherichia coli into
a transgenic
bacterium
INDUSTRIAL INSULIN PRODUCTION
• The transgenic
bacteria are then
selected and
cultured in a
fermentation tank to
increase bacterial
numbers
INDUSTRIAL INSULIN PRODUCTION
• The bacteria now
expresses the insulin
gene and produce
human insulin, which
is harvested, purified
and packaged for
human use i.e. by
diabetics
ANTICODON
Understanding
Translation depends on complementary base pairing
between codons on mRNA and anticodons on tRNA
ANTICODON
•It is a
sequence of
three bases
on an tRNA
molecule
TRANSLATION
Understanding
Translation is the synthesis of polypeptides on
ribosomes
TRANSLATION
•Translation is
the synthesis
of
polypeptides
by ribosomes
TRANSLATION
• Translation is the
process of protein
synthesis in which the
genetic information
encoded in mRNA is
translated into a
sequence of amino acids
on a polypeptide chain
TRANSLATION
•Each codon of
three bases on the
mRNA is translated
into one amino acid
in a polypeptide
chain
TRANSLATION
• Additionally, the
sequence of the
codons on the
mRNA determines
the amino acid
sequence of the
polypeptide made
TRANSLATION
• Translation involves
the following
components
• mRNA
• Ribosomes
• tRNA
THE MECHANISM OF
TRANSLATION
Understanding
Translation depends on complementary base pairing
between codons on mRNA and anticodons on tRNA
THE MECHANISM OF TRANSLATION
• Ribosomes bind to
mRNA in the
cytoplasm and move
along the molecule in
a 5’ – 3’ direction until
it reaches a start
codon (AUG)
THE MECHANISM OF TRANSLATION
• Anticodons on tRNA
molecules align
opposite appropriate
codons according to
complementary base
pairing (e.g. AUG =
UAC)
THE MECHANISM OF TRANSLATION
•Each tRNA
molecule carries
a specific amino
acid (according
to the genetic
code)
THE MECHANISM OF TRANSLATION
• Ribosomes catalyse
the formation of
peptide bonds
between adjacent
amino acids (via
condensation
reactions)
THE MECHANISM OF TRANSLATION
• The ribosome
moves along the
mRNA molecule
synthesising a
polypeptide chain
until it reaches a
stop codon
THE MECHANISM OF TRANSLATION
•At this point
translation
ceases and the
polypeptide
chain is
released
POLYMERASE CHAIN
REACTION (PCR)
Application
Use of Taq DNA polymerase to produce multiple copies of
DNA rapidly by the polymerase chain reaction (PCR)
POLYMERASE CHAIN REACTION (PCR)
• The polymerase chain
reaction, generally
known as PCR, is a
method for rapid
production of a very
large number of copies
of a particular fragment
of DNA
POLYMERASE CHAIN REACTION (PCR)
• PCR is a technique
that can make billions
of copies of one
molecule of DNA by
repeatedly copying a
specific stretch of that
DNA
POLYMERASE CHAIN REACTION (PCR)
• The PCR technique
for amplifying DNA
was developed by
Kary Mullis in 1983,
earning him a Nobel
prize for his work
POLYMERASE CHAIN REACTION (PCR)
• The technique uses
the cyclic heating and
cooling of a DNA
sample in the presence
of primers, DNA
nucleotides and Taq
polymerase to amplify
the DNA
POLYMERASE CHAIN REACTION (PCR)
• Taq polymerase is
a DNA polymerase
isolated from a
bacterium,
Thermus aquaticus
which lives in hot water
springs at temperatures
between 50°C and 80°C
POLYMERASE CHAIN REACTION (PCR)
• As this enzyme’s
optimal temperature
is ~75ºC, it is able to
function at the high
temperatures used in
PCR without
denaturing
STEPS INVOLVED IN PCR
APPLICATIONS OF PCR
Application
Use of Taq DNA polymerase to produce multiple copies of
DNA rapidly by the polymerase chain reaction (PCR)
APPLICATIONS OF PCR
• PCR has enabled
scientists to clone
genes, to work with
minute amounts of
DNA found at crime
scenes and identify
the dead
APPLICATIONS OF PCR
•It can also be
used to sequence
the DNA of extinct
humans and other
life forms
BIBLIOGRAPHY / ACKNOWLEDGMENTS
END OF UNIT