16/10/2023, 08:34
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Evidence for semi-conservative replication-Meselson and Stahl’s experiment
Meselson and Stahl showed DNA is replicated using the semi-conservative
method. Their experiment used two isotopes of nitrogen (DNA contains nitrogen)
— heavy nitrogen (15N) and light nitrogen (14N).
This experiment was based on 3 important facts
1. All DNA havses contain nitrogen(14N)
2. Nitrogen has 2 forms, lighter nitrogen and heavy nitrogen (15N
3. Babestia will incorporate nitrogen from their browth medium into the new
DNA rhwy will make
1. Two samples of bacteria were grown for many generations — one in a nutrient
broth containing light nitrogen, and one in a broth with heavy nitrogen. As the
bacteria reproduced, they took up nitrogen from the broth to help make
nucleotides for new DNA. So the nitrogen gradually became part of the bacteria’s
DNA.
2. A sample of DNA was taken from each batch of bacteria, and spun in a centrifuge.
The DNA from the heavy nitrogen bacteria settled lower down the centrifuge tube
than the DNA from the light nitrogen bacteria — because it’s heavier
3. Then the bacteria grown in the heavy nitrogen broth were taken out and put in a
broth containing only light nitrogen. The bacteria were left for one round of DNA
replication, and then another DNA sample was taken out and spun in the
centrifuge
4. If replication was conservative, the original heavy DNA, which would still be
together, would settle at the bottom and the new light DNA would settle at the top
5. If replication was semi-conservative, the new bacterial DNA molecules would
contain one strand of the old DNA containing heavy nitrogen and one strand of
new DNA containing light nitrogen. So the DNA would settle out between where
the light nitrogen DNA settled out and where the heavy nitrogen DNA settled out
6. As it turned out, the DNA settled out in the middle, showing that the DNA
molecules contained a mixture of heavy and light nitrogen. The bacterial DNA had
replicated semi-conservatively in the light nitrogen.
Once Meselson and Stahl had confirmed that DNA replication in bacteria was
semi-conservative, other scientists carried out experiments to show that it was the
universal method for DNA replication in all living things.
print.
How is DNA replicated?
A DNA molecule has a paired base structure , which makes it easy for DNA to
copy itself. Here’s how it works:
1. The enzyme DNA helicase breaks the hydrogen bonds between bases on the two
polynucleotide DNA strands (complemenatry bases pairsbe between a double
helix)). This makes the helix unwind to form two single strands
The action of DNA polymerase
Each end of a DNA strand is slightly different in its structure. One end is called
the 3’ (pronounced ‘three prime’) end and one end is called the 5’ (five prime)
end.
During DNA replication the active site of DNA polymerase is only
complementary to the 3’ end of the newly forming DNA strand— so the enzyme
can only add nucleotides to the new strand at the 3’ end.
This means that the new strand is made in a 5’ to 3’ direction and that DNA
polymerase moves down the template strand in a 3’ to 5’ direction
.
2. Each original single strand acts as a template for a new strand. Complementary
base pairing means that free-floating DNA nucleotides are attracted to their
complementary exposed bases on each original template strand — A with T and C
The 3’ and 5’ ends of a DNA strand are different. At the 3’ end is a hydroxyl group
(OH) that is attached to the pentose sugar of the nucleotide. At the 5’ end there is a
phosphate group. Take a look back at page 54 for a reminder of the structure of
DNA.
Because the strands in the double helix are antiparallel, the DNA polymerase
working on one of the template strands moves in the opposite direction to the
with G. DNA polymerase working on the other template strand
3. The adjacent nucleotides are joined together (to form the phosphodiester bonds by
a condensation reaction).DNA polymerase catalyses the joining together of
adjacent nucleotides
4. Condensation reactions join the nucleotides of the new strand together —
catalysed by the enzyme DNA polymerase. Hydrogen bonds form between the
bases on the original and new strands.
Each new DNA molecule contains one strand from the original DNA molecule
and one new strand.
The 2 sets of daughter DNA (name of the new DNA molecules)contain one strand
of the parental (original ) DNA and one newly synthesised strand
16/10/2023, 08:34 print.html

Why does DNA replicate?

DNA copies itself before cell division so that each new cell has the full amount of
DNA.
The method is called semi-conservative replication because half of the strands in
each new DNA molecule are from the original DNA molecule.
This means that there’s genetic continuity between generations of cells (i.e. the
cells produced by cell division inherit their genes from their parent cells)