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25/07/22

DNA Replication
State why DNA replication is semi-conservative.
Nucleotide Monomers &
ATP
Describe the process of DNA replication.
DNA Structure &
Modelling
Explain the roles of the enzymes helicase and DNA polymerase
in DNA replication. DNA Purification (PAG 9)

Replication
Starter:
Theories & PEQs

The Genetic Code

THINK: Why does DNA need to replicate?
Transcription & Translation

Summary
 H A R E
S Why does DNA need to replicate?
1) DNA copies itself before cell division so that each new
cell has the full amount of DNA.
2) This is important for:
a) Making new cells for growth and repair.
b) For passing genetic information from generation to
generation (reproduction).
3) The DNA replicate must be an exact copy to form two
sister chromatids, and so the process must be accurate.
State why DNA replication is semi-conservative.
Describe the process of DNA replication
In pairs, use the images to summarise DNA replication.

Keywords to use:
Unzips, hydrogen bonds, DNA
helicase, free nucleotides,
complementary, DNA
polymerase, identical, semi-
DNA polymerase
conservative. 10
minutes

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Describe the process of DNA replication
1. The enzyme DNA helicase breaks the hydrogen bonds
2. The double helix separates and the two strands unwind
3. Each strand asks as a template, complementary free nucleotides bind to their
specific base on the template
4. Nucleotides are joined together in a condensation reaction by DNA polymerase
5. Each new DNA molecule contains one of the original strand and one newly
synthesised strand – semi conservative replication

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 H
WATC Roles of Enzymes in Replication

DNA replication is controlled by enzymes, a class of proteins that

act as catalysts for biochemical reactions.

While watching the video by

Amoeba Sisters on DNA
replication, give the roles of
helicase and DNA polymerase.

Explain the roles of the enzymes helicase and DNA polymerase in DNA replication.
 E
SHAR Helicase
KEY TERM

Helicase: Enzyme that

catalyses the breaking of
hydrogen bonds between
the nitrogenous pairs of
bases in a DNA molecule.

The result is two single

strands of DNA with
exposed nucleotide bases.

Explain the roles of the enzymes helicase and DNA polymerase in DNA replication.
 E
SHAR DNA Polymerase
KEY TERM

DNA polymerase:
Enzyme that catalyses
the formation of DNA
from activated
deoxyribose nucleotides,
using single stranded
DNA as a template.
Catalyses the formation of phosphodiester bonds between the
nucleotides.
This addition of nucleotide bases happens in the 5’ to 3’ direction.
Explain the roles of the enzymes helicase and DNA polymerase in DNA replication.
 E
SHAR Enzymes involved in DNA replication

TIP The role of the enzyme polymerase is to attach to

one end of the nucleotide chain and covalently bond other
nucleotides to the chain to form the polynucleotide. As
enzymes are specific, there are two types; DNA polymerase
and RNA polymerase.
Explain the roles of the enzymes helicase and DNA polymerase in DNA replication.
 E Process of DNA Replication
SHAR

EXAM TIP

If answering an exam question

about semi-conservative
replication, you need to make sure
you do it in the correct order, or
you won’t get all of the marks.

Describe the process of DNA replication.

 H
MATC Enzymes involved in DNA replication
DNA ligase Joins together short sections of the lagging
strand.

Helicase Catalyses the formation of a new

polynucleotide chain.

DNA polymerase Keeps the separated DNA strands apart during

replication.

Single-strand Separates the two DNA strands before

binding protein replication.

Explain the roles of the enzymes helicase and DNA polymerase in DNA replication.
 E Process of DNA Replication
SHAR
3) Free nucleotides join the
1) DNA helicase 2) Double helix complementary unpaired bases.
causes hydrogen structure
bonds to break, unwinds.
separating strands.

5) DNA
4) Hydrogen bonds polymerase
form between bases. catalyses the
formation of
phosphodiester
bonds between
Half the molecules is old DNA, half is the nucleotides.
made of new molecules. This is known
as semi-conservative replication.
Describe the process of DNA replication.
Two theories existed...
Conservative Hypothesis
The complete parent DNA molecule acts as a template
for the new daughter molecule, which is assembled from
new nucleotides. The parent molecule is unchanged.

Semi-conservative Hypothesis
The parent DNA molecule separates into its two
component strands, each of which acts as a template for
the formation of a new complementary strand. The two
daughter molecules therefore contain half the parent
DNA and half new DNA (semi-conservative hypothesis).
 E
SHAR Semi-conservative replication
The product of the replication is two DNA molecules, identical to each other
and to the parent molecule.

Each of these molecules contains one old strand and one new strand, as so is
termed semi-conservative replication.
The loops of DNA in prokaryotes, and KEY TERM
inside mitochondria and chloroplasts,
also replicate semi-conservatively. A
Semi-conservative replication: Each new
bubble sprouts from the loop and this DNA molecule is formed of one conserved
unwinds and unzips. The complementary strand from the original molecule, which
nucleotides join to the exposed acts as the template, and one strand of
nucleotides. Eventually the whole loop is new nucleotides.
copied.
State why DNA replication is semi-conservative.
 E Continuous and Discontinuous Replication
SHAR
1) DNA polymerase always moves along the template strand in the same direction. It can only
bind to the 3’ (OH) end, so travels in the direction of 3’ to 5’.
2) As DNA only unwinds and unzips in one direction, DNA polymerase has to replicate each of
the template strands in opposite directions.
3) The strand that is unzipped from the 3’ end can be continuously replicated. This strand is
called the leading strand and is said to undergo continuous replication.
4) The other strand is unzipped from the 5’
end, so DNA polymerase has to wait until a
section of the strand has been unzipped and
then work back along.
5) This results in DNA being produced in
sections, called Okazaki fragments. These
fragments then have to be joined. This strand
is called the lagging strand and is said to
undergo discontinuous replication.
Describe the process of DNA replication.
Lets draw it out
 TASK Process of DNA Replication

1) Complete the summary of DNA replication by

reordering the statements and creating a poster to
show the processes.

2) Draw a diagram to show semi-conservative

replication.
Describe the process of DNA replication.
 S
ASSES
1) Both strands act as template.
2) This is called the semi-conservative theory of DNA replication.
3) H bonds between base pairs break and the DNA unzips.
4) Hydrogen bonds reform between base pairs.
5) DNA untwists by the action of DNA helicase
6) The sugar-phosphate back bone forms, joining by a phosphodiester
bond.
7) This is catalysed by DNA polymerase.
8) Free nucleotides align to the DNA and complementary base pair, C to
G and T to A.
9) Two new DNA molecules form, each with one parent strand and one
new strand.
Describe the process of DNA replication.
 S
ASSES
1) DNA untwists by the action of DNA helicase.
2) H bonds between base pairs break and the DNA unzips.
3) Both strands act as template.
4) Free nucleotides align to the DNA and complementary base pair, C to
G and T to A.
5) Hydrogen bonds reform between base pairs.
6) The sugar-phosphate back bone forms, joining by a phosphodiester
bond.
7) This is catalysed by DNA polymerase.
8) Two new DNA molecules form, each with one parent strand and one
new strand.
9) This is called the semi-conservative theory of DNA replication.
Describe the process of DNA replication.
THE MOST BEAUTIFUL EXPERIMENT I
N BIOLOGY: Meselson & Stahl, The Semi-C
•onservative Replication
The semi conservative ofshown
hypothesis was DNA - the
to be YouTube
true
mechanism by the work of Meselsohn and Stahl (1958).
Meselsohn and Stahl (1958)
• They grew the bacteria Escherichia coli with different isotopes of
Nitrogen.
• The bacteria was exposed to N15 for several generations until it was
exposed to a lighter N14.
• Scientists could then distinguish between the different DNA densities
by centrifuging them.
They knew...
• All the bases in DNA contain Nitrogen

• Nitrogen has two forms:

• Light 14N
• Heavy isotope 15N

• Bacteria will incorporate nitrogen from their growing medium into

any new DNA they make
Proving replication is
semi-conservative
•The 15N strand is heavy so makes a
band low down the tube
•High density (heavy) sinks further
down the tube.

After one generation

•After one generation there was only
one band
•This was the 14/15 hybrid

Second generation
•After another generation there were 2
bands
•A light chain and a 14/15 level
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Complete the questions
Answer the Past Exam Questions.
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Self Assess
1. Table 5.1 contains a list of statements about DNA replication. Some of these statements
are incorrect. Put a cross (X) in the box next to each incorrect statement. [3]
Incorrect
DNA replication
statement
The DNA molecule unwinds  
Hydrogen bonds between the base pairs break  
Free RNA nucleotides join to bases on the exposed DNA strands X 
Both polypeptide strands act as a template X 
Hydrogen bonds form between complementary bases  
Three hydrogen bonds form between bases A and T X 
DNA polymerase links the new nucleotides  
Covalent bonds form between the phosphate of one nucleotide  
and the pentose sugar of the next nucleotide
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Self Assess
2.A gene codes for the production of lactase. This gene is normally switched off
after an infant moves to adult food. Almost all adult mammals are unable to
digest lactose. They are said to be lactose intolerant. Humans are an exception.
 
Most humans have a genetic mutation that prevents the shutdown of lactase
production.
 
State what structural detail of a polypeptide is altered by gene mutations. [1]

Sequence/order of amino acids (Allow primary structure)

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Self Assess
3.i. Outline how the process of DNA replication is completed,
following the pairing of nitrogenous bases. [3]
(involves) DNA polymerase (1)
sugar-phosphate backbone (re)forms / condensation
reaction between phosphate and sugar (1)
DNA winds into double helix (1)
ii. Why is DNA replication described as semi-conservative? [1]
(new molecule consists of) one old strand and one new
strand (1)
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Self Assess
4. Semi-conservative replication describes the process by which DNA is replicated in all living
organisms.
i. Explain the meaning of the phrase semi-conservative replication. [1]
(new DNA molecule comprises)
one, original / old / parent, strand
and one new strand ✔
each strand (of DNA molecule) acts as a template strand
(for a new double helix) ✔
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Self Assess
4.
ii. DNA ligase is one enzyme involved in the replication of DNA.
State two other enzymes involved and describe their functions. [4]
E1 (DNA) helicase ✔
F1 unzips the DNA molecule / breaks hydrogen bonds (between complementary
bases) / separates the (2) strands ✔
E2 DNA polymerase ✔
F2 forms phosphodiester bonds / joins (adjacent) nucleotides / forms sugar-
phosphate backbone ✔
Also creditworthy
E3 gyrase ✔
F3 unwinds / uncoils, the DNA ✔
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Self Assess
5. The chromosomes carried by sperm are made of DNA.

The following passage about nucleic acids has four words missing. Choose the correct missing
words from the list below and complete the passage by writing them in the gaps.

nucleotide monomers.
Nucleic acids are made from ...................................

phosphate
Phosphodiester bonds form between the monomers. They consist of a ...................................
pentose
group between the ................................... molecules, forming the ‘backbone’ of the molecule.

strands
In DNA, hydrogen bonding between the two antiparallel ................................... causes the
characteristic double helix shape. [4]
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Self Assess
6. DNA replication and transcription are two processes that occur in the nucleus of
eukaryotic cells.* Compare DNA replication and transcription by describing the similarities
and differences between the two processes. [6]
Similarities Differences
• DNA unwinds and unzips • Only a small section of DNA (where the gene is
• Helicase enzymes located) unzips during transcription
• Template DNA • Both strands act as templates in replication
• Complementary base • RNA vs DNA free nucleotides
pairing • RNA vs DNA polymerase
• Hydrogen bonds • Different helicase enzymes
• Free, activated • Products are two new daughter strands of DNA
nucleotides in replication and one mRNA strand in transcription
• Polymerase enzymes • mRNA leaves nucleus whereas the new DNA
strand remains bound to the template strand
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Self Assess
7. A group of students attempted to extract and purify DNA from a plant in Upper End Meadow.
The students used the following steps:
1. Mix the plant sample with detergent. 2. Add salt.
3. Add protease enzyme. 4. Spool the DNA precipitate onto a glass rod.
Suggest whether this method would successfully extract and purify DNA. Justify your conclusion. [3]
Y1 detergent, breaks / disrupts, (cell) membrane(s) / nuclear envelope OR detergent,
releases contents of, cell / nucleus ✓
Y2 salt, helps DNA, shed water / precipitate ✓
Y3 protease breaks down, histones / proteins around DNA / proteins attached to DNA
✓

NO reasons why it would not, work / be successful: 2 max

N1 cell walls not broken by, abrasion / grinding / blender ✓
N2 no RNAase added to remove RNA (from DNA / chromatin) ✓
N3 no, alcohol / ethanol, added to, precipitate DNA ✓
N4 temperature not low to reduce, enzyme activity / DNA break down ✓