Mol Bio Da1

DIGITAL ASSIGNMENT -1
1. In Griffith’s experiment what was called “Transforming principle” and

why this name was given?
The transforming principle was an early name for DNA. In 1928, scientists didn't
know yet that DNA carried genetic information, but they knew that there was
something that could cause bacteria to transform from one type to another.
Bacteriologist Frederick Griffith (in 1928) 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.
• 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.
• 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 nonvirulent,
meaning that they did not cause sickness when injected into a mouse.
1. Mice infected with the S strain (virulent) die from pneumonia infection.
S Strain ------------> Inject into mice--------------->Mice died
2. Mice infected with the R strain do not develop pneumonia.
R Strain ------------> Inject into mice--------------->Mice lived
3. Griffith was able to kill bacteria by heating them. He observed that heat-killed S
strain bacteria injected into mice did not kill them.
S Strain (heat-killed) ------------> Inject into mice--------------->Mice lived
4. When he injected a mixture of heat-killed S and live R bacteria, the mice died.
Moreover, he recovered living S bacteria from the dead mice.
S Strain (heat-killed) + R Strain (live)-------> Inject into mice-------->Mice died
He concluded that the R strain bacteria had somehow been transformed by the heat-
killed S strain bacteria. Some ‘transforming principle’, transferred from the heat-killed
S strain, had enabled the R strain to synthesise a smooth polysaccharide coat and
become virulent. This must be due to the transfer of the genetic material.
However, the biochemical nature of genetic material was not defined from his
experiments.
So genetic material was called transforming principle .
2. What did Avery, MacLeod and McCarty proved in their experiment?

Why scientists were not convinced that the DNA is the transforming
principle even in this experiment?
In 1944, three Canadian and American researchers, Oswald Avery, Maclyn McCarty,
and Colin MacLeod, set out to identify Griffith's "transforming principle."
They took large cultures of heat-killed S cells and, through a long series of
biochemical steps , progressively purified the expected transforming principles(
DNA, RNA and PROTEIN) by washing away, separating out, or enzymatically
destroying the other cellular components.
By this method, they were able to obtain small amounts of highly purified
transforming principle, which they could then analyze through other tests to
determine its identity.
Several lines of evidence suggested to Avery 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- degrading enzymes (Protease) and RNA-degrading
enzymes(RNases) had little effect on the transforming principle, but enzymes
able to degrade DNA (DNases) eliminated the transforming activity.
These results all pointed to DNA as the likely transforming principle.
However, many scientist and biologist were still not convinced that DNA is the
transforming principle because they thought it is still possible that some
contaminating substance present in small amounts, not DNA, was the actual
transforming principle
Because of this possibility, debate over DNA's role continued until 1952, when Alfred
Hershey and Martha Chase used a different approach to conclusively identify DNA
as the genetic material.
3. Why Hershey and Chase experiment also called as blender experiment?
What purpose the blender was used in these experiments?
In 1952, Alfred Hershey, a Carnegie Institution researcher working at Cold Spring

Harbor Laboratory, proved DNA as genetic material. Like many of his
contemporaries, Hershey believed that proteins, with their complicated structures,
were more likely to be the carriers of genetic information than was the simple DNA
molecule.
In order to show that proteins carry genetic information, Hershey and his lab
technician, Martha Chase, decided to track the transfer of proteins and DNA
between a virus and its host. For their experiment, they chose to use the T2
bacteriophage as the vehicle for delivering genetic material. Like all bacterial
viruses, the T2 is comprised of only a protein-based outer wall and a DNA core, its
simple structure making it the perfect research candidate. The phage reproduces by
injecting its genetic material into a bacterium, leaving its protein shell attached to the
host. Then, through a microscopic takeover, the virus seizes control of the
bacterium’s reproductive mechanisms and uses them to duplicate itself, destroying
the host in the process.
In their experiment, to establish whether the phage injected DNA or protein into host
bacteria, Hershey and Chase:-
1. Prepared two different batches of phage. In each batch, the phage 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 sample was produced in the presence of Sulphur(35), a radioactive isotope
of sulphur. Sulphur is found in many proteins and is absent from DNA, so only
phage proteins were radioactively labelled by this treatment.
• The other sample was produced in the presence of Phosphorus(32), a
radioactive isotope of phosphorous. Phosphorous is found in DNA and not in
proteins, so only phage DNA (and not phage proteins) was radioactively
labelled by this treatment.
Each batch of phage was used to infect a different culture of bacteria.
3. After infection had taken place, each culture was whirled in a blender, removing
any remaining phage and phage parts from the outside of the bacterial cells.
4. Finally, the cultures were centrifuged, or spun at high speeds, to separate the
bacteria from the phage debris.
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 P(32) appeared in the pellet , indicating that DNA was the material
that passed from the virus to the bacteria.
Whereas almost all of the S(32) appeared in the supernatant, indicating that proteins
did not enter the bacteria from the viruses.
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.
Blender used in the experiment for removing any phage or phage parts from the
outside of bacterial cell, therefore the experiment also called the blender experiment

Mol Bio Da1

Uploaded by

Copyright:

Available Formats

Mol Bio Da1

Uploaded by

Document Information

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Mol Bio Da1

Uploaded by

Copyright:

Available Formats

DIGITAL ASSIGNMENT -1

1. In Griffith’s experiment what was called “Transforming principle” and

2. What did Avery, MacLeod and McCarty proved in their experiment?

In 1952, Alfred Hershey, a Carnegie Institution researcher working at Cold Spring

You might also like

Pfad - The Proxy pFad of © 2024 Garber Painting. All rights reserved.