Mbt01- Cell Bio-wps Office

MBT01- CELL BIOLOGY
Topic- Replication of DNA, semi-conservative mode of replication
Contents:
1. Introduction
2. Replication of dna
3. Meaning of semi-conservative model
4. Meselson and stahl experiment
5. Models of replication:
6.Separation and recombination of double-stranded DNA
7.Conclusion
1
Introduction:
Semiconservative replication describe the mechanism of DNA replication in all

known cells. DNA replication occurs on multiple origins of replication along the
DNA template strands. As the DNA double helix is unwound by helicase,
replication occurs separately on each template strand in antiparallel directions.
This process is known as semi-conservative replication because two copies of the
original DNA molecule are produced, each copy conserving (replicating) the
information from one half of the original DNA molecule. Each copy contains one
original strand and one newly synthesized strand. (Both copies should be
identical, but this is not entirely assured.) The structure of DNA (as deciphered by
James D. Watson and Francis Crick in 1953) suggested that each strand of the
double helix would serve as a template for synthesis of a new strand. It was not
known how newly synthesized strands combined with template strands to form
two double helical DNA molecules.
2.DNA replication:
In the process of DNA replication, the DNA makes multiple copies of itself. It is a
biological polymerisation, which proceeds in the sequence of initiation,
elongation, and termination. It is an enzyme-catalysed reaction. DNA Polymerase
is the main enzyme in the replication process.
DNA Replication Steps
2
Following are the important steps involved in DNA replication:
1.Initiation
DNA replication demands a high degree of accuracy because even a minute

mistake would result in mutations. Thus, replication cannot initiate randomly at
any point in DNA.
For the replication to begin there is a particular region called the origin of
replication. This is the point where the replication originates. Replication begins
with the spotting of this origin followed by the unwinding of the two DNA strands.
Unzipping of DNA strands in their entire length is not feasible due to high energy
input. Hence, first, a replication fork is created catalysed by the helicase enzyme,
which unzips the DNA strand.
2.Elongation
As the strands are separated, the polymerase enzymes start synthesising the
complementary sequence in each of the strands. The parental strands will act as a
template for newly synthesising daughter strands.
It is to be noted that elongation is unidirectional i.e. DNA is always polymerised

only in the 5′ to 3′ direction. Therefore, in one strand (the template 3‘→5‘) it is
continuous, hence called continuous replication while on the other strand (the
template 5‘→3‘) it is discontinuous replication. They occur as fragments called
Okazaki fragments. The enzyme called DNA ligase joins them later.
3.Termination
Termination of replication occurs in different ways in different organisms. In E.coli

like organisms, chromosomes are circular. And this happens when the two
replication forks between the two terminals meet each other.
3.Meaning of semi-conservative model:

Semi-conservative replication means that during DNA replication, the two strands of
nucleotides separate. Both strands then form the template for free nucleotides to bind to to
3
create the two identical daughter strands. Hence each daughter strand has half of the DNA
from the original strand and half newly-formed DNA.
4.Meleson and Stahl experiment:

Multiple experiments were conducted to determine how DNA replicates. The semiconservative
model was anticipated by Nikolai Koltsov and later supported by the Meselson–Stahl
experiment, which confirmed that DNA replicated semi-conservatively by conducting an
experiment using two isotopes: nitrogen-15 (15N) and nitrogen-14 (14N). When 14N was added
to the heavy 15. that experiment done on E-coli bacterium in culture medium.
N-15N DNA, a hybrid of 15N-14N was seen in the first generation. After the second generation,
the hybrid remained, but light DNA (14N-14N) was seen as well. This indicated that DNA
replicated semi-conservatively. This mode of DNA replication allowed for each daughter strand
to remain associated with its template strand.
4
5.Models of replication:
5
Semiconservative replication derives its name from the fact that this mechanism of
transcription was one of three models originally proposed for DNA replication:
Semiconservative replication would produce two copies that each contained one of the original
strands of DNA and one new strand. Semiconservative replication is beneficial to DNA repair.
During replication, the new strand of DNA adjusts to the modifications made on the template
strand.
Conservative replication would leave the two original template DNA strands together in a
double helix and would produce a copy composed of two new strands containing all of the new
DNA base pairs.
Dispersive replication would produce two copies of the DNA, both containing distinct regions
of DNA composed of either both original strands or both new strands. The strands of DNA were
originally thought to be broken at every tenth base pair to add the new DNA template.
Eventually, all new DNA would make up the double helix after many generations of replication.
6.Separation and recombination of double-stranded DNA

For semiconservative replication to occur, the DNA double-helix needs to be separated so the
new template strand can be bound to the complementary base pairs. Topoisomerase is the
enzyme that aids in the unzipping and recombination of the double-helix. Specifically,
topoisomerase prevents the double-helix from supercoiling, or becoming too tightly wound.
Three topoisomerase enzymes are involved in this process: Type IA Topoisomerase, Type IB
Topoisomerase, and Type II Topoisomerase. Type I Topoisomerase unwinds double stranded
DNA while Type II Topoisomerase breaks the hydrogen bonds linking the complementary base
pairs of DNA.
6.Applications:
*Semiconservative replication provides many advantages for DNA. It is fast, accurate, and
allows for easy repair of DNA. It is also responsible for phenotypic diversity in a few prokaryotic
species.
*The process of creating a newly synthesized strand from the template strand allows for the old
strand to be methylated at a separate time from the new strand. This allows repair enzymes to
proofread the new strand and correct any mutations or errors.
6
*DNA could have the ability to activate or deactivate certain areas on the newly synthesized
strand that allows the phenotype of the cell to be changed. This could be advantageous for the
cell because DNA could activate a more favorable phenotype to aid in survival.
*Due to natural selection, the more favorable phenotype would persist throughout the species.
This gives rise to the idea of inheritance, or why certain phenotypes are inherited over another.
7. Conclusion:
The experiment done by Meselson and Stahl demonstrated that DNA replicated semi-
conservatively, meaning that each strand in a DNA molecule serves as a template for synthesis
of a new, complementary strand. Although Meselson and Stahl did their experiments in the
bacterium E.coli. Semi-conservative mode of replication produces two copies, each containing
one original strand and one new strand. On the contrary, conservative replication produces two
new strands and would leave two original template DNA strands in a double helix.

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MBT01- CELL BIOLOGY

Topic- Replication of DNA, semi-conservative mode of replication

3. Meaning of semi-conservative model

4. Meselson and stahl experiment

6.Separation and recombination of double-stranded DNA

Semiconservative replication describe the mechanism of DNA replication in all

DNA Replication Steps

DNA replication demands a high degree of accuracy because even a minute

It is to be noted that elongation is unidirectional i.e. DNA is always polymerised

Termination of replication occurs in different ways in different organisms. In E.coli

3.Meaning of semi-conservative model:

4.Meleson and Stahl experiment:

6.Separation and recombination of double-stranded DNA

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