Topic D1.1.

Meselson's and Stahl Model

They cultured E. coli bacteria in the presence of a heavy nitrogen isotope, 15 N. DNA contains
nitrogen in its nitrogenous bases (ATCG), so the radioactive 15 N would end up in the DNA of
the bacteria. The result was that all bacterial DNA had 15 N in its bases.
They then transferred the bacterial culture into a fresh medium where the nitrogen was
replaced by 14 N, a lighter isotope, and the bacteria were allowed to grow for several
generations.
DNA samples were then extracted from successive bacterial generations and subjected to
caesium chloride equilibrium density gradient centrifugation. This technique allowed the
DNA to move to different positions in the centrifuge tube based on its density. DNA
containing one or two strands with 15 N was heavier and showed lower bands than those
containing two strands with 14 N (DNA with 15 N in both strands was heaviest).

Figure 2. Results obtained from the Meselson and Stahl experiment.

After one generation, i.e., one division of the bacteria, the resulting DNA strand consisted of
a double helix, where one strand was made up of 15 N, and the other contained 14 N. The band
obtained was in between those for DNA with both strands containing 15 N or 14 N only. This
indirectly demonstrated that DNA replication had to proceed in a semi-conservative way and
should involve complementary base pairing to ensure the fidelity of the daughter molecules.
Results of the successive generations of bacteria further confirmed this theory.

Note: each of two or more forms of the same element that contain equal numbers of protons but
different numbers of neutrons in their nuclei, and hence differ in relative atomic mass but not in
chemical properties; in particular, a radioactive form of an element.

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Topic D1.1. Meselson's and Stahl Model

1. Bacteria cultured in a medium containing only 15N were transferred to a medium

containing only 14N and allowed to complete one round of replication. The DNA in bacteria
produced because of replication on the 14N medium was extracted and subjected to cesium
chloride centrifugation which separates DNA molecules according to their density.

Which centrifuge tube shows the arrangement of bands observed after one round of
replication?

A. Tube 3

B. Tube 4

C. Tube 5

D. Tube 6

2. For what did Meselson and Stahl’s work provide evidence?

A. The abiotic origin of organic molecules

B. The cell theory

C. The fluid mosaic model of membrane structure

D. The semi-conservative replication of DNA

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Topic D1.1. Meselson's and Stahl Model

3. Meselson and Stahl investigated DNA replication by first feeding bacteria with bases
containing 15N (heavy), and then with bases containing 14N (light). The results are shown in
the photographs.

From this information, what would be the composition of the DNA in generation 3?

A. There will be a single 14N band.

B. There will be a single 15N band and a single 14N band.

C. There will be a darker 14N band and a lighter hybrid band.

D. There will be a single 15N band, a single hybrid band and a single 14N band.

4. Meselson and Stahl conducted experiments using the isotopes 14N and 15N which showed
that DNA replication is semi-conservative. What would they have observed about the
distribution of isotopes in the DNA after one round of replication if DNA replication was
conservative rather than semi-conservative?

A. Only 14N DNA

B. Only 15N DNA

C. All DNA half 14N and half 15N

D. Half the DNA with only 14N and half with only 15N

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Topic D1.1. Meselson's and Stahl Model

5. Cells were grown in heavy nitrogen (15N) for many generations and then grown in light
nitrogen (14N) for two rounds of DNA replication. Which diagram shows the result of the
centrifuged DNA?

6. Which are necessary to make DNA replication semi-conservative?

I. Separation of the strands by RNA polymerase

II. Complementary base pairing

III. Use of a pre-existing strand as a template

A. I and II only

B. I and III only

C. II and III only

D. I, II and III

FUN ACTIVITY

The following strand of DNA needs to be replicated. You are to complete this process
by adding the correct bases in sequence using the complementary base pairing rule. It
is a race, however, so you need to time yourself to see how fast you can do it.

ATCCGTATCAATGGCAATGCCCTAGATGCCGTTAGACCTTTTAGAGCA-
CATA