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DNA As Genetic Material

- Griffith conducted experiments with two strains of bacteria, R and S, finding that R bacteria could be transformed into virulent S bacteria by exposure to heat-killed S bacteria, indicating a "transforming principle". - Avery, McCarty, and MacLeod sought to identify the transforming principle and through chemical and enzymatic tests determined it was likely DNA. - Hershey and Chase used bacteriophage labeled with radioactive isotopes of phosphorus or sulfur to track DNA and protein during viral infection, finding that DNA, not protein, entered host cells and must be the genetic material.

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72 views19 pages

DNA As Genetic Material

- Griffith conducted experiments with two strains of bacteria, R and S, finding that R bacteria could be transformed into virulent S bacteria by exposure to heat-killed S bacteria, indicating a "transforming principle". - Avery, McCarty, and MacLeod sought to identify the transforming principle and through chemical and enzymatic tests determined it was likely DNA. - Hershey and Chase used bacteriophage labeled with radioactive isotopes of phosphorus or sulfur to track DNA and protein during viral infection, finding that DNA, not protein, entered host cells and must be the genetic material.

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maghfoorkhakh247
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DNA as genetic

material
Course instructor: Fatima Sadiq
• Proteins were known to have diverse amino acid
sequences, while DNA was thought to be a boring,
repetitive polymer, due to an incorrect (but popular)
model of its structure and composition.
• Today, we know that DNA is not actually repetitive
and can carry large amounts of information. But how
did scientists first come to realize that "boring" DNA
might actually be the genetic material?
Frederick Griffith: Bacterial
transformation
• In 1928, British bacteriologist Frederick Griffith
conducted a series of experiments
using Streptococcus pneumoniae bacteria and mice.
Griffith wasn't trying to identify the genetic material,
but rather, trying to develop a vaccine against
pneumonia. In his experiments, Griffith used two
related strains of bacteria, known as R and S.
• R strain. When grown in a petri dish, the R bacteria
formed colonies, or clumps of related bacteria, that
had well-defined edges and a rough appearance
(hence the abbreviation "R"). The R bacteria were
non-virulent, meaning that they did not cause
sickness when injected into a mouse.
• S strain. S bacteria formed colonies that were
rounded and smooth (hence the abbreviation
"S"). The smooth appearance was due to a
polysaccharide or sugar-based, coat produced
by the bacteria. This coat protected the S
bacteria from the mouse immune system,
making them virulent (capable of causing
disease). Mice injected with live S bacteria
developed pneumonia and died.
• As part of his experiments, Griffith tried injecting
mice with heat-killed S bacteria (that is, S bacteria
that had been heated to high temperatures, causing
the cells to die). Unsurprisingly, the heat-killed S
bacteria did not cause disease in mice.
• Griffith concluded that the R-strain bacteria must
have taken up what he called a "transforming
principle" from the heat-killed S bacteria, which
allowed them to "transform" into smooth-coated
bacteria and become virulent.
Avery, McCarty, and MacLeod:
Identifying the transforming principle

• In 1944, three Canadian and American researchers,


Oswald Avery, Maclyn McCarty, and Colin MacLeod,
set out to identify Griffith's "transforming principle."
• Several lines of evidence suggested to Avery and his
colleagues that the transforming principle might be
DNA.
• The purified substance gave a negative result in
chemical tests known to detect proteins, but a strongly
positive result in a chemical test known to detect DNA.
• The elemental composition of the purified
transforming principle closely resembled DNA in its
ratio of nitrogen and phosphorous.
• Protein- and RNA-degrading enzymes had little effect
on the transforming principle, but enzymes able to
degrade DNA eliminated the transforming activity.
• These results all pointed to DNA as the likely
transforming principle. However, Avery was cautious
in interpreting his results. He realized that it was still
possible that some contaminating substance present in
small amounts, not DNA, was the actual transforming
principle.
The Hershey-Chase experiments

• In their now-legendary experiments, Hershey and


Chase studied bacteriophage, or viruses that attack
bacteria. The phages they used were simple particles
composed of protein and DNA, with the outer
structures made of protein and the inner core
consisting of DNA.
• Hershey and Chase knew that the phages attached to
the surface of a host bacterial cell and injected some
substance (either DNA or protein) into the host. This
substance gave "instructions" that caused the host
bacterium to start making lots and lots of phages—in
other words, it was the phage's genetic material.
Before the experiment, Hershey thought that the
genetic material would prove to be protein
• To establish whether the phage injected DNA or
protein into host bacteria, Hershey and Chase
prepared two different batches of phage. In each
batch, the phages were produced in the presence of a
specific radioactive element, which was incorporated
into the macromolecules (DNA and protein) that
made up the phage
• One batch of phage was labeled with 35S, which is
incorporated into the protein coat. Another batch was
labeled with 32P, which is incorporated into the DNA.
• Bacteria were infected with the phage.
• The cultures were blended and centrifuged to separate
the phage from the bacteria.
• Radioactivity was measured in the pellet and liquid
(supernatant) for each experiment. 32P was found in
the pellet (inside the bacteria), while 35S was found in
the supernatant (outside of the bacteria)
• Centrifugation causes heavier material, such as
bacteria, to move to the bottom of the tube and form a
lump called a pellet. Lighter material, such as the
medium (broth) used to grow the cultures, along with
phage and phage parts, remains near the top of the
tube and forms a liquid layer called the supernatant
• When Hershey and Chase measured radioactivity in
the pellet and supernatant from both of their
experiments, they found that a large amount of P32,
appeared in the pellet, whereas almost all of the S35
appeared in the supernatant. Based on this and
similar experiments, Hershey and Chase concluded
that DNA, not protein, was injected into host cells
and made up the genetic material of the phage.

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