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CHAPTER 6 Exp Notes Class 12

Alfred Hershey and Martha Chase conducted experiments in 1952 that provided unequivocal proof that DNA is the genetic material. They cultured bacteriophages in media containing radioactive phosphorus or sulfur to label the phage DNA or proteins. When the labeled phages infected E. coli bacteria, only the bacteria infected with phosphorus-labeled phages (DNA) became radioactive, demonstrating that DNA enters the bacteria during infection. Their conclusion was that DNA is the genetic material passed from viruses to bacteria.

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199 views

CHAPTER 6 Exp Notes Class 12

Alfred Hershey and Martha Chase conducted experiments in 1952 that provided unequivocal proof that DNA is the genetic material. They cultured bacteriophages in media containing radioactive phosphorus or sulfur to label the phage DNA or proteins. When the labeled phages infected E. coli bacteria, only the bacteria infected with phosphorus-labeled phages (DNA) became radioactive, demonstrating that DNA enters the bacteria during infection. Their conclusion was that DNA is the genetic material passed from viruses to bacteria.

Uploaded by

Meeta Biswas
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© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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CHAPTER 6

NOTES

EXPERIMENTS OF ALFRED HERSHEY AND MARTHA CHASE


(1952).

DNA as Genetic Material


 The unequivocal proof that DNA is the genetic material came from the
experiments of Alfred Hershey and Martha Chase (1952).
 They worked with viruses that infect bacteria called bacteriophages.
 The bacteriophage attaches to the bacteria and its genetic material then
enters the bacterial cell.
 The bacterial cell treats the viral genetic material as if it was its own and
subsequently manufactures more virus particles.
 Hershey and Chase worked to discover whether it was protein or DNA
from the viruses that entered the bacteria.

Experiment: The experiment began with the culturing of viruses in


two types of medium.
 One set of viruses (A) was cultured in a medium of radioactive
phosphorus
 Whereas another set (B) was cultured in a medium of radioactive
sulfur.
 They observed that the first set of viruses (A) consisted of
radioactive DNA but not radioactive proteins.
 This is because DNA is a phosphorus-based compound while
protein is not.
 The latter set of viruses (B) consisted of radioactive protein but
not radioactive DNA.
 The host for infection was E.coli bacteria.
 The viruses were allowed to infect bacteria by removing the viral
coats through a number of blending and centrifugation.
OBSERVATION:
 E.coli bacteria which were infected by radioactive DNA viruses
(A) were radioactive
 But the ones that were infected by radioactive protein viruses
(B) were non-radioactive.

CONCLUSION:
 Resultant radioactive and non-radioactive bacteria infer that
the viruses that had radioactive DNA transferred their DNA to
the bacteria
 The viruses that had radioactive protein didn’t get transferred
to the bacteria.
 Bacteria that were infected with viruses that had radioactive proteins
were not radioactive. This indicates that proteins did not enter the
bacteria from the viruses.
 DNA is therefore the genetic material that is passed from virus to
bacteria

MESELSON-STAHL EXPERIMENT
 This experiment was performed to prove the SEMI
CONSERVATIVE NATURE of DNA replication.
 Matthew Meselson & Franklin Stahl experimented with
bacteria E.coli in 1958.

EXPERIMENT
15
1. N (heavy) and 14N (normal) are two isotopes of nitrogen, which
can be distinguished based on their densities by centrifugation in
Caesium chloride (CsCl).
2. Meselson and Stahl cultured E.coli in a medium
constituting 15NH4Cl over many generations. As a result, 15N was
integrated into the bacterial DNA.
3. They grew E. coli in a medium containing 15NH4Cl (15N is the heavy
isotope of nitrogen) as the only nitrogen source for many generations.
4. The result was that 15N was incorporated into newly synthesised DNA (as
well as other nitrogen containing compounds).
5. This heavy DNA molecule could be distinguished from the normal DNA
by centrifugation in a cesium chloride (CsCl) density gradient ( 15N is not
a radioactive isotope, and it can be separated from 14N only based on
densities).
6. Then they transferred the cells into a medium with normal 14NH4Cl and
took samples at various definite time intervals as the cells multiplied,
and extracted the DNA that remained as double-stranded helices.
7. The various samples were separated independently on CsCl gradients to
measure the densities of DNA .

OBSERVATIONS :
 Thus, the DNA that was extracted from the culture one generation after
the transfer from 15N to 14N medium [that is after 20 minutes; E. coli
divides in 20 minutes] had a hybrid or intermediate density.
 DNA extracted from the culture after another generation [that is after 40
minutes, II generation] was composed of equal amounts of this hybrid
DNA and of ‘light’ DNA.

CONCLUSION :
 Based on observations and experimental results, Meselson and
Stahl concluded that DNA molecules can replicate semi-
conservatively.

CBSE QUESTIONS :
Q 1 Answer the following questions based on Hershey and Chases’s
experiments

(a) Name the kind of virus they worked with and why ?

(b) Why did they use two types of culture media to grow viruses in ? Explain.

(c) What was the need for using a blender and later a centrifuge during their
experiments ?

(d) State the conclusion drawn by them after the experiments.

ANSWERS :
(a) They worked with bacteriophage which infect bacteria because they want
to discover whether it was protein or DNA from the viruses that entered the
bacteria.

(b) They used two types of culture media in order to make protein of viruses
radioactive with the help of 35S in one case, and DNA molecule in virus
radioactive by using 32P in the other case. This was done to identify which
one of the two had entered into the bacteria during viral infection.

(c) Blender was used to separate viral protein coats that – were still attached
to the surface of bacteria.
Centrifuge was used to separate lighter supernatent containing viral protein
coats from denser residue containing bacteria.

(d) They concluded that DNA is the genetic material that is passed from virus
to bacteria.

Q2 (a) What did Meselson and Stahl observe when


(i) They cultured E. coil in medium containing 15NH4Cl for a few
generations and centrifuged the content.
(ii) They transferred one such bacterium to the normal medium of NH 4Cl
and cultured for 2 generations?
(b) What did Meselson and Stahl conclude from this experiment? Explain
with the help of diagrams.
(c) Which is the first genetic material? Give reasons in support of your
answer.
(a)

i.) Meselson and Stahl observed that in the E. coli bacterium the DNA
becomes completely labelled with N medium by centrifugation for few
generations.
ii.) After two generations, density changed and showed equal amount of
light DNA (N14) and dark hybrid DNA (N–N).
(b) (b) They concluded that DNA replicates semi-conservatively.
c )Ribonucleic acid (RNA) was the first genetic material.

Q.3 : State the aim describe Messelson and Stahl experiment. 5 mks
Q 4 : Name the scientist who proved that DNA is the genetic material.
5 marks
Q. 5 Describe the experiment that helped demonstrate the semi-
conservative mode of DNA replication. – 5Marks.

ANSWER :

SEMI-CONSERVATIVE DNA REPLICATION:


The two Strands would separate and act as a template for the
synthesis of new complementary strands. After the completion of
replication, each DNA molecule would have one parental and one
newly sytnthesized strads this scheme was termed as semi-
conservative DNA replication.

Who Found: In 1958 Meselson and Stahl's proved the semi-


conservative mode of replication.

Experiment:

1. They worked with E. coli using a heavy isotope of Nitrogen, 15N


E. coli was grown with 15N for several generation till the
bacteria become completely labelled with 15N.

2. They were then shifted to medium with normal 14N nitrogen


sample were taken after every genereation and tested for the
presence of 15N and 14N using DENSITY GRADIANT
CENTRIFUGATION with CsCI .
3. The first generation was found be hybrid between 15N and 14N
second generation contain two types of DNA, - 50% light (14N)
and 50% intermediate.

4. This is only possible if two strands separate during replication


and act as a template for synthesis of new complementary
strands, thus proving a semiconservative mode of replication.
OBSERVATION:

 Since E. coli divides every 20 minutes, the DNA extracted after


20 minutes experiment had a hybrid density.
 The DNA extracted after 40 minutes had equal amounts of
hybrid and light densities.

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