From the Specification

DNA replication
The semi-conservative replication of DNA ensures genetic continuity
between generations of cells.​
The process of semi-conservative replication of DNA in terms of:​
•unwinding of the double helix​
•breakage of hydrogen bonds between complementary bases in the
polynucleotide strands​
•the role of DNA helicase in unwinding DNA and breaking its hydrogen
bonds​
•attraction of new DNA nucleotides to exposed bases on template strands
and base pairing​
•the role of DNA polymerase in the condensation reaction that joins
adjacent nucleotides.​
Students should be able to evaluate the work of scientists in validating the
Watson–Crick model of DNA replication.
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C/W
How is DNA copied to form 2 Monday 10 June

identical molecules?
2024

Learning Outcomes:
• Compare and contrast the structure of DNA and RNA
• Describe the process of semi-conservative replication of DNA
• Predict results from experiments used to prove that DNA
replication is semi-conservative

Starter Activity:

Describe the structure of DNA

Draw a single DNA nucleotide

Homework
DNA Questions
One Nucleotide
RNA
 RNA is single stranded and relatively
short.

Monday, June 10, 2024 Explain the structure of RNA 7

 ATGCCTGAAACGTACAGT

What is the complementary RNA sequence?

Monday, June 10, 2024 8

Explain the role of RNA

Three types of RNA:

1. RNA that transfers genetic information from DNA to
ribosomes (mRNA).
2. RNA that makes up ribosomes along with protein (rRNA).
3. RNA involved in protein synthesis (tRNA).

Monday, June 10, 2024 9

1. RNA that transfers genetic information from DNA to
ribosomes (mRNA).

First step to making

proteins.

We will learn this later.

Monday, June 10, 2024 Explain the role of RNA 10

2. RNA that makes up ribosomes along with protein (rRNA).

rRNA and protein make up ribosomes.

Ribosomes make proteins.

Monday, June 10, 2024 Explain the role of RNA 11

3. RNA involved in protein synthesis (tRNA).

tRNA binds to mRNA to make a chain

of amino acids (protein).

Monday, June 10, 2024 Explain the role of RNA 12

Compare and contrast the similarities and differences between
DNA and RNA
DNA RNA
Name
Chain length
Strand number
Pentose sugar
Organic bases
Chemical stability
Function

Monday, June 10, 2024 13

Compare and contrast the similarities and differences between
DNA and RNA
DNA RNA
Deoxyribonucleic acid Ribonucleic acid
Name
Very long Relatively short
Chain length
Double Single
Strand number
Deoxyribose Ribose
Pentose sugar
Thymine, guanine, cytosine Uracil, guanine, cytosine and
Organic bases and adenine adenine
Very stable Less stable
Chemical stability
Carries genetic information Transfer information from
DNA to ribosomes. Make up
ribosomes with protein.
Function Involved in protein
synthesis.

Monday, June 10, 2024 14

The genetic code – important things to remember!
A gene is a sequence of bases which code for making polypeptides.

Each code has three bases- a triplet code (e.g. TAC). Some amino acids
only have a single triplet code – others have 2-6.

How many amino acids does the following sequence of bases code
for? TACGCTCCGCTGTAC

There are 20 amino acids, but there are 64 different codes possible!

Some amino acids have more than one code. This is called the
degenerate code.
In eukaryotes much of the nuclear DNA does not code for amino acids
– sometimes called junk DNA (98% with a largely unknown function !)
Mini - Plenary: What am I? On mini-whiteboards
• ‘I’m made up a phosphate group, ribose sugar and
organic base’
• ‘I contain three bases and code for an amino acid’
• ‘I’m found on chromosomes and code for polypeptides’
• ‘We hold together base pairings in a DNA molecule, this
means we hold the two strands together'
• ‘There are 20 varieties of me’
• ‘I'm a chemical reaction - when I form bonds between
nucleotides a molecule of water is released’
• The name of the bonds formed between two
nucleotides
 The Importance of DNA Replication

• When cells divide the daughter cell must receive an

exact copy of the genetic material from the parent.

• In order for this to happen the DNA must be

replicated or copied.
1. DNA helicase unzips the 2 strands.
What is DNA helicase?

The enzymes break the hydrogen bonds holding the base pairs together.
2. Free nucleotides bind
Complementary base pairing

• DNA helicase completes the

splitting of the strand.
• Meanwhile, free nucleotides
are attracted to their
complementary bases.
• Each chain acts as a template.
• RNA segments act as primers
Why might this be known as
semi-conservative replication?

Each new DNA double helix contains one new strand and one old strand
3. DNA Polymerase bonds neighbouring
nucleotides together
• Once in placed the activated
nucleotides are joined
together by DNA
polymerase.
• DNA polymerase joins the
new nucleotides to each
other by strong covalent
bonds, forming the
phosphate-sugar backbone.

What are the bonds that are

formed called?

Phosphodiester bonds
 The final result
The result is that there are two DNA molecules, each with one new
synthesised strand of DNA and one strand from the original.
The two strands in DNA are anti-parallel
What does this mean?
https://www.youtube.com/watch?v=pz3vZ7HDnvQ
https://www.youtube.com/watch?v=NGLuO-NYRug
https://www.youtube.com/watch?v=qWZYpHSXvJo
https://www.youtube.com/watch?v=0CQ5ls3Uc2Q
 Written Work
1. Describe how DNA is copied during semi-
conservative replication.
2. Summarise the role of the following enzymes:
• DNA helicase
• DNA polymerase
• (DNA ligase)
 Steps of Replication

1. Replication starts at a specific sequence on the DNA molecule.

2. DNA helicase unwinds and unzips DNA, breaking the hydrogen bonds
that join the base pairs, and forming two separate strands.
3. The new DNA is built up from the four nucleotides (A, C, G and T)
that are abundant in the nucleoplasm.
4. These nucleotides attach themselves to the bases on the old strands
by complementary base pairing. Where there is a T base, only an A
nucleotide will bind, and so on.
5. DNA polymerase joins the new nucleotides to each other by strong
covalent bonds, forming the phosphate-sugar backbone.
6. A winding enzyme winds the new strands up to form double helices.
7. The two new molecules are identical to the old molecule.
Explain what 5' and 3' ends mean in regards to DNA structure?
5' and 3' ends describes the directionality of the DNA molecule. Essentially, the strand of
a DNA molecule can have a 5' end and a 3' end. To understand what a 5' or 3' end is, we
need to look at the molecular structure of DNA. DNA is a polymer of nucleotides, where
each nucleotide is made up of a sugar (deoxyribose), a nitrogenous base, and a
phosphate group. The deoxyribose sugar is a 5 carbon structure, where each carbon can
be numbered 1-5. The base is always connected to Carbon 1 of the sugar and the
phosphate group is connected to Carbon 5 of the sugar. The nucleotides are then
connected to one another to form the polymer whereby the phosphate group of one
nucleotide (on Carbon 5) connects to the next nucleotide sugar via Carbon 3. Therefore
when the strand is built, the top nucleotide will have a free phosphate group on the
Carbon 5 of the sugar, hence the 5' end, and the last nucleotide of the strand will have a
free OH on the Carbon 3 of the sugar. Since DNA is made up of two antiparallel strands,
each strand can have its own directionality. As they are antiparallel, they will run in
opposite directions.
 Give two features of DNA and explain how each
one is important in the semi-conservative
replication of DNA. (2 marks)

1. Weak / easily broken hydrogen bonds between bases

allow two strands to separate / unzip;
2. Two strands, so both can act as templates;
3. Complementary base pairing allows accurate
replication;
Semi-Conservative Model
 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).
2 models
 Evidence for the
semi-conservative model
• The semi conservative hypothesis was shown to be the 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.

Predict what will happen when the mixture is centrifuged

again.

What would have happened if the conservative model was

correct?
Proving
replication is
semi-
conservative
After one generation
After one generation
there was only one
band
It was higher than the
heavy DNA band
This was the 14/15
hybrid.

Predict what will happen when the mixture is grwon for

another generation and then centrifuged again.​
 Proving replication
is semi-conservative

Second generation
•After another
generation there
were 2 bands
•A light chain and a
14/15 level chain
Exam Questions
1. Give two features of DNA and explain how each one is important
in the semi-conservative replication of DNA. (2)
2. Replication of mitochondrial DNA (mtDNA) is different from that of
nuclear DNA. The replication of the second strand of mtDNA only
starts after two-thirds of the first strand of mtDNA has been
copied. A piece of mtDNA is 16 500 base pairs long and is
replicated at a rate of 50 nucleotides per second. (1)
3. A scientist replicated DNA in a test tube. To do this, he mixed an
enzyme with identical single-stranded DNA fragments and a
solution containing DNA nucleotides.
(a) Name the enzyme used in this DNA replication. (1)
(b) Use your knowledge of semi-conservative replication of DNA to
suggest:
1. the role of the single-stranded DNA fragments
2. the role of the DNA nucleotides. (3)
Answers
(1)
1. Weak / easily broken hydrogen bonds between bases allow two strands to separate /
unzip;
may appear in the same feature
2. Two strands, so both can act as templates;
may appear in the same feature
3. Complementary base pairing allows accurate replication;
Allow description of complementary base pairing and accurate replication.
2 max
(2) 550 seconds;
1
(3) (a) DNA polymerase;
Correct spelling
1
(b) Role of single-stranded DNA fragments
1. Template;
2. Determines order of nucleotides/bases;
Role of DNA nucleotides
3. Forms complementary pairs / A – T, G - C
OR
Forms complementary (DNA) strand;
Ignore forms complementary bases 3