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2/11/2020 Mode of DNA replication: Meselson-Stahl experiment (article) | Khan Academy

Science · Biology · DNA as the gene c material


· DNA replica on

Mode of DNA replica on:


Meselson-Stahl experiment
A key historical experiment that demonstrated the semi-
conserva ve mechanism of DNA replica on.

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Key points:
There were three models for how organisms
might replicate their DNA: semi-conserva ve,
conserva ve, and dispersive.

The semi-conserva ve model, in which each


strand of DNA serves as a template to make a
new, complementary strand, seemed most
likely based on DNA's structure.

The models were tested by Meselson and Stahl,


who labeled the DNA of bacteria across
genera ons using isotopes of nitrogen.

From the pa erns of DNA labeling they saw,


Meselson and Stahl confirmed that DNA is
replicated semi-conserva vely.

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Mode of DNA replica on


Imagine yourself in 1953, a er the double helix
structure of DNA has just been discovered1 . What
burning ques ons might be on your mind, and on
the minds of other scien sts?

One big ques on concerned DNA replica on. The


structure of the DNA double helix provided a
tantalizing hint about how copying might take
place1,2 . It seemed likely that the two
complementary strands of the helix might separate
during replica on, each serving as a template for
the construc on of a new, matching strand.

But was this actually the case? Spoiler alert: The


answer is yes! In this ar cle, we'll look at a famous
experiment, some mes called "the most beau ful
experiment in biology," that established the basic
mechanism of DNA replica on as semi-
conserva ve—that is, as producing DNA molecules
containing one new and one old strand3 .

The three models for DNA


replica on
There were three basic models for DNA replica on
that had been proposed by the scien fic
community a er the discovery of DNA's structure.
These models are illustrated in the diagram below:

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_Image modified from "Basics of DNA replica on: Figure 1," by OpenStax
College, Biology (CC BY 3.0)._

Semi-conserva ve replica on. In this model,


the two strands of DNA unwind from each
other, and each acts as a template for synthesis
of a new, complementary strand. This results in
Science Biology DNA as
two DNA molecules with one original strand
the gene c material DNA
replica on and one new strand.
DNA replica on

Leading and lagging


Conserva ve replica on. In this model, DNA
strands in DNA replica on replica on results in one molecule that consists
of both original DNA strands (iden cal to the
Speed and precision of
DNA replica on original DNA molecule) and another molecule

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Molecular structure of that consists of two new strands (with exactly


DNA
the same sequences as the original molecule).
Molecular mechanism of
DNA replica on Dispersive replica on. In the dispersive model,
DNA replica on results in two DNA molecules
Mode of DNA replica on: that are mixtures, or “hybrids,” of parental and
Meselson-Stahl
experiment daughter DNA. In this model, each individual
strand is a patchwork of original and new DNA.
DNA proofreading and
repair
Most biologists at the me would likely have put
Telomeres and telomerase
their money on the semi-conserva ve model. This
model made a lot of sense given the structure of
Prac ce: DNA replica on the DNA double helix, in which the two DNA
strands are perfectly, predictably complementary to
one another (where one has a T, the other has an A;
where one has a G, the other has a C; and so
forth)2,4 . This rela onship made it easy to imagine
each strand ac ng as a template for the synthesis
of a new partner.

However, biology is also full of examples in which


the “obvious” solu on turns out not to be the
correct one. (Protein as the gene c material,
anyone?). So, it was key to experimentally
determine which model was actually used by cells
when they replicated their DNA.

Meselson and Stahl cracked the


puzzle

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Ma Meselson and Franklin Stahl originally met in


the summer of 1954, the year a er Watson and
Crick published their paper on the structure of
DNA. Although the two researchers had different
research interests, they became intrigued by the
ques on of DNA replica on and decided to team
up and take a crack at determining the replica on
mechanism5 .

The Meselson-Stahl experiment


Meselson and Stahl conducted their famous
experiments on DNA replica on using E. coli
bacteria as a model system.

They began by growing E. coli in medium, or


nutrient broth, containing a "heavy" isotope of
nitrogen, 15 N. (An isotope is just a version of an
element that differs from other versions by the
number of neutrons in its nucleus.) When grown on
medium containing heavy 15 N, the bacteria took up
the nitrogen and used it to synthesize new
biological molecules, including DNA.

A er many genera ons growing in the 15 N


medium, the nitrogenous bases of the bacteria's
DNA were all labeled with heavy 15 N. Then, the
bacteria were switched to medium containing a
"light" 14 N isotope and allowed to grow for several
genera ons. DNA made a er the switch would

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have to be made up of 14 N, as this would have been


the only nitrogen available for DNA synthesis.

Meselson and Stahl knew how o en E. coli cells


divided, so they were able to collect small samples
in each genera on and extract and purify the DNA.
They then measured the density of the DNA (and,
indirectly, its 15 N and 14 N content) using density
gradient centrifuga on.

This method separates molecules such as DNA into


bands by spinning them at high speeds in the
presence of another molecule, such as cesium
chloride, that forms a density gradient from the top
to the bo om of the spinning tube. Density
gradient centrifuga on allows very small
differences—like those between 15 N- and 14 N-
labeled DNA—to be detected.

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_Image modified from "Meselson-Stahl experiment diagram en," by


Mariana Ruiz Villareal (public domain)._

Results of the experiment


When DNA from the first four genera ons of E. coli
was analyzed, it produced the pa ern of bands
shown in the figure below:

_Image modified from "Basics of DNA replica on: Figure 2," by OpenStax
College, Biology (CC BY 3.0). Original artwork from "Meselson-Stahl
experiment diagram en," by Mariana Ruiz Villareal (public domain)._

What did this result tell Meselson and Stahl? Let's


walk through the first few genera ons, which
provide the key informa on.

Genera on 0
DNA isolated from cells at the start of the
experiment (“genera on 0,” just before the switch
to 14 N medium) produced a single band a er
centrifuga on. This result made sense because the
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DNA should have contained only heavy 15 N at that


me.

Genera on 1
DNA isolated a er one genera on (one round of
DNA replica on) also produced a single band when
centrifuged. However, this band was higher,
intermediate in density between the heavy 15 N
DNA and the light 14 N DNA.

The intermediate band told Meselson and Stahl


that the DNA molecules made in the first round of
replica on was a hybrid of light and heavy DNA.
This result fit with the dispersive and semi-
conserva ve models, but not with the conserva ve
model.

The conserva ve model would have predicted two


dis nct bands in this genera on (a band for the
heavy original molecule and a band for the light,
newly made molecule).

Genera on 2
Informa on from the second genera on let
Meselson and Stahl determine which of the
remaining models (semi-conserva ve or dispersive)
was actually correct.

When second-genera on DNA was centrifuged, it


produced two bands. One was in the same posi on
as the intermediate band from the first genera on,
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while the second was higher (appeared to be


labeled only with 14 N).

This result told Meselson and Stahl that the DNA


was being replicated semi-conserva vely. The
pa ern of two dis nct bands—one at the posi on
of a hybrid molecule and one at the posi on of a
light molecule—is just what we'd expect for semi-
conserva ve replica on (as illustrated in the
diagram below). In contrast, in dispersive
replica on, all the molecules should have bits of old
and new DNA, making it impossible to get a "purely
light" molecule.

_Image modified from "Basics of DNA replica on: Figure 2," by OpenStax
College, Biology (CC BY 3.0). Original artwork from "Meselson-Stahl
experiment diagram en," by Mariana Ruiz Villareal (public domain)._

Genera ons 3 and 4


In the semi-conserva ve model, each hybrid DNA
molecule from the second genera on would be
expected to give rise to a hybrid molecule and a
light molecule in the third genera on, while each
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light DNA molecule would only yield more light


molecules.

Thus, over the third and fourth genera ons, we'd


expect the hybrid band to become progressively
fainter (because it would represent a smaller
frac on of the total DNA) and the light band to
become progressively stronger (because it would
represent a larger frac on).

As we can see in the figure, Meselson and Stahl


saw just this pa ern in their results, confirming a
semi-conserva ve replica on model.

[What if the dispersive model had been correct?]


[What if the conserva ve model had been correct?]

Conclusion
The experiment done by Meselson and Stahl
demonstrated that DNA replicated semi-
conserva vely, 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, we know today that semi-
conserva ve DNA replica on is a universal
mechanism shared by all organisms on planet Earth.
Some of your cells are replica ng their DNA semi-
conserva vely right now!
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[A ribu on and references]

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Shailu 3 years ago


more

Why is Cesium chloride used? Why can't the


centrifuga on be done without it?
(14
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lea sha.phillips 4 years ago


more

what causes the double helix of the DNA to


"unzip"
(6
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4 years
m…
isaacboardman ago

During DNA replica on, the enzyme


helicase unwinds the DNA double
helix by disrup ng the hydrogen bonds
that keep it together. Different
proteins are also involved in the
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unzipping of the double helix such as


single strand binding proteins that
keep the two strands from reforming
hydrogen bonds.
2 (17
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viegasmaya 2 years ago


more

Could someone explain to me the results of the


3rd and 4th genera on please? I don't quite
understand that part, why isn't all DNA semi-
conserva ve? Why is there a N^14 light strand?
(3
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Taka Tanabe 2 years ago


more

1st we know: Bacteria is grown in N15


,
then placed in N14 .
(remember N14 is the light isotope
and N15 is the heavy one.)
So ini ally all nitrogenous bases of
each nucleo de will have contained
N15 isotope.
So once place in N14 medium ,DNA
replica on will use the N14 Isotope
only.
No more N15 available.
Thats why in the 3rd and 4th
genera on , the N15 strand disappears
over me
So what remains are DNA Strands that
undergo Semi conserva ve replica on
with the fixed amount of N15 and an
increasing amount of N14, as N14 is
available in the medium.
The chart in Fig. 2 kind of explains it.
(6
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Comment p m
votes)

Adi.price 4 years ago


more

when its spli ng is there every a case when it


doesn't split correctly
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Emily 4 years ago


more

Absolutely! There are numerous


syndromes, diseases and condi ons
based on the improper spli ng. This is
something that is easily googled and
you can find tons of informa on on!
(6
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Julie S 2 years ago


more

What would have happened if they grew the


bacteria in the light isotope and then introduced
it to the heavy isotope? What would the vial
layers and densi es look like? (Essen ally, what
would happen if you did the opposite of what
they did.)
(4
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7

months
Ivana - Science trainee ago

I think as for results experiment would


be the same. Giving the same results.
Density gradient would always display
N15 on the bo om and N14 above it,
there is no ques on.

Not sure about the cost. Introducing


heavy isotope later or earlier makes no

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difference in cost if they have to use


both.
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1024tom27 4 years ago


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what is A, T, G, C in DNA Helix


2 (0
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4 years
m…
Todor Cvetanovic ago

Those are purines and pyrimidines,


those are the nitrogen bases that build
DNA molecule. A stands for Adenine,
G stands for Guanine, and those two
are purines. And pyrimidines are C that
stands for Cytosine and T stands for
Thymine.
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Bonin Sok 2 years ago


more

If it is concluded that each strand serves as a


template for the synthesis of a new
complementary strand, how come a hybrid
strand does not produce two hybrid molecules.
For example, one light and one heavy template
strand, produces a complementary heavy and
light strand respec vely.
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Jen 2 years ago


more

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Good ques on! Keep in mind that the


DNA is grown in N15 (heavy) but the
next genera ons are in N14 (light). So
a hybrid strand cannot produce hybrid
molecules because there aren't N15-
labelled nucleo des available! The
ques on and response below might
help clarify too :)
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Hannah a month ago


more

Why are bacteria like Escherichia coli good for


studying DNA replica on (as opposed to using
other types of cells)?
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a
m…
month
Ivana - Science trainee ago

E. coli is model organism that's why.

Why E. coli model organism?


E. coli is a perfect model for several
reasons: E. coli is a single-celled
organism that can be manipulated and
killed with no ethical concerns. It has a
rapid growth rate and is very easy to
culture and grow.

h ps://study.com/academy/lesson/esc
herichia-coli-e-coli-as-a-model-
organism-or-host-cell.html

The main reasons why E. coli is the


organism of choice extends and is not
limited to its fast growth in chemically
defined media; rela ve cheap culture
media; does not form aggregates;

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industrial scalability; several molecular


tools for manipula on; extensive
knowledge of its gene cs and
genomics; extensive knowledge on its
transcriptome, proteome, and
metabolome, and several strains are
considered biosafety 1, which renders
it ideal even for teaching and school
demonstra ons.

In terms of ecology, E. coli is a


faculta ve aerobe (either respira on
takes place in the presence of oxygen
or fermenta on in its absence), which
bears a sensor for oxygen presence
(redox state in the quinone pool) and
can ac vate or repress the required
metabolic enzymes, depending on
oxygen levels.

The building blocks of E. coli consists


of about 55% protein, 25% nucleic
acids, 9% lipids, 6% cell wall, 2.5%
glycogen, and 3% other metabolites
[16–18], which for biotechnological
applica ons is important since carbon
flux is o en a problema c issue to
address to generate a novel metabolic
pathway or to enhance a current
func oning pathway.

E. coli is part of the normal microbiota


of mammals, rendering the
predominant faculta ve microbe of
the gastrointes nal tract and is
currently a hot debate on the impact
on normal microflora establishment
and their role in disease.

E. coli harbours a genome with


par cular features such as a strikingly
organized structure, remnants of many
phages, and inser on sequences (IS)
and a high transport capacity toward

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the cytoplasm.

h ps://www.intechopen.com/books/-
i-escherichia-coli-i-recent-advances-
on-physiology-pathogenesis-and-
biotechnological-applica ons/-i-
escherichia-coli-i-as-a-model-
organism-and-its-applica on-in-
biotechnology
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Darya Behrooz a year ago


more

Would the results be different if they didn't use


E. Coli but something else instead?
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7

months
Ivana - Science trainee ago

No, because DNA replicates semi-


conserva vely. :)

Maybe the experiment would be set


up slightly different but results must
be the same.

Replica on is semi-conserva ve in
both: Eukaryotes and Prokaryotes
meaning that it is the same for any
living organism.
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Sundus a year ago


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It means the other two models don't exist?


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tyersome a year ago


more

Models are proposed explana ons for


or descrip ons of something.

All three of the models discussed in


this ar cle "exist", but one of them is a
be er descrip on of how DNA
replica on actually occurs.

Does that help?


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Show more...

Molecular mechanism of DNA replica on


DNA proofreading and repair

Biology is brought to you with support from the

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