DNA as Genetic Material

Direct Evidences
 IDENTIFICATION OF DNA AS
THE GENETIC MATERIAL
• The genetic material must meet 4 criteria
– 1. Information:
– 2. Transmission:
– 3. Replication:
– 4. Variation:

9-3
Search for the genetic material:

1. Stable source of information

2. Ability to replicate accurately
3. Capable of change

Timeline of events:

• 1890 Weismann - substance in the cell nuclei controls development.

• 1900 Chromosomes shown to contain hereditary information,

later shown to be composed of protein & nucleic acids.

• 1928 Griffith’s Transformation Experiment (incorrectly guessed protein!)

• 1944 Avery’s Transformation Experiment (DNA not RNA)

• 1953 Hershey-Chase Bacteriophage Experiment (DNA not protein)

• 1956 First demonstration that RNA is viral genetic material.

Bacterial Transformation
Bacterial Transformation
 Avery’s Experiments
 Mixed each extract with type IIR cells to test for transformation
 Only extract containing purified DNA transformed type IIR to type IIIS
 Verify that DNA, not RNA or protein, is the genetic material

Figure 9.3 9-11

 1600s 1800s 1850s 1900s 1950s 2000s

1877-1955
• Oswald Avery – American bacteriologist

S R

DNA

S
– 1943 – proved that DNA carries genes
 Griffith’s transformation experiment

Peter J. Russell, iGenetics: Copyright © Pearson Education, Inc., publishing as Benjamin Cummings.
 Experiment that showed that DNA, not RNA, was the transforming principle

Peter J. Russell, iGenetics: Copyright © Pearson Education, Inc., publishing as Benjamin Cummings.
 Avery, MacLeod, McCarty experiment

Fig. 6.4 c
 Bacterial Transduction

• In 1952, Hershey and Chase, studying

bacteriophages (T2), labelled DNA with 32P and
protein with 35S
• Bacteriophage progeny produced by infection
of bacteria contained 32P (thus DNA from the
original phage), but not 35S (from the protein)!
Hershey-Chase Bacteriophage Experiment - 1953

Bacteriophage = Virus that attacks

bacteria and replicates by
invading a living cell and using
the cell’s molecular machinery.

Fig. 2.4
Structure of T2 phage

Bacteriophages
are composed of
DNA & protein
DNA discovery Hershey-Chase 1952
Fig. 2.5: Life cycle of virulent T2 phage:
Fig. 2.6: Hershey-Chase Bacteriophage Experiment - 1953

1. T2 bacteriophage is composed
of DNA and proteins:

2. Set-up two replicates:

• Label DNA with 32P

• Label Protein with 35S

3. Infected E. coli bacteria with

two types of labeled T2

4. 32P is discovered within the

bacteria and progeny phages,
whereas 35S is not found within
the bacteria but released with
phage ghosts.

Alfred Hershey
 Bacterial Conjugation

• In 1946, Lederberg and Tatum showed that

two different strains of bacteria with different
growth requirements could exchange genes
• Lederberg and Tatum surmised that the
bacterial cells must interact with each other -
the process is now known as sexual
conjugation
Joshua Lederberg and Edward Tatum,
studied two strains of Escherichia coli with
different nutritional requirements. Strain

A would grow on a minimal medium only

if the medium were supplemented with
methionine and biotin; strain B would
grow on a minimal medium only if it were
supplemented with threonine, leucine,
and thiamine. Thus, we can designate
strain A as met− bio− thr+ leu+ thi+ and
strain B as met+ bio+ thr− leu− thi−.

Strains A and B are mixed together, and

some of the progeny are now wild type,
having regained the ability to grow
without added nutrients
It could be suggested that the cells of the
two strains do not really exchange genes
but instead leak substances that the other
cells can absorb and use for growing.

This possibility of “cross feeding” was ruled

out by Bernard Davis.
He constructed a U-tube in which the two
arms were separated by a fine filter. The
pores of the filter were too small to allow
bacteria to pass through but large enough
to allow easy passage of the fluid medium
and any dissolved substances.
Strain A was put in one arm; strain B in the
other. After the strains had been incubated
for a while, Davis tested the content of
each arm to see if cells had become able to
grow on a minimal medium, and none were
found. In other words, physical contact
between the two strains was needed for
wild-type cells to form. It looked as though
some kind of gene transfer had taken place,
and genetic recombinants were indeed
produced.
Transfer of F plasmid
The F plasmid directs the synthesis of pili, projections that initiate contact with a recipient
and draw it closer, allowing the F DNA to pass through a pore into the recipient cell. One
strand of the double-stranded F DNA is transferred and then DNA replication restores the
complementary strand in both the donor and the recipient.
RNA As Genetic Material
Fig. 2.7 (2nd edition)
Gierer & Schramm Tobacco Mosaic Virus (TMV) Experiment - 1956
Fraenkel-Conrat & Singer - 1957

Demonstrated that RNA is the genetic material of TMV.

Conclusions about these early experiments:

Griffith 1928 & Avery 1944:

DNA (not RNA) is transforming agent.

Hershey-Chase 1953:

DNA (not protein) is the genetic material.

Gierer & Schramm 1956/Fraenkel-Conrat & Singer 1957:

RNA (not protein) is genetic material of some viruses, but no known

prokaryotes or eukaryotes use RNA as their genetic material.

Alfred Hershey
Nobel Prize in Physiology or Medicine
1969
INDIRECT EVIDENCES OF DNA AS GENETIC
MATERIAL
Bacteria, viruses and other microorganisms show
direct evidences to prove that DNA is the genetic material but
higher organisms show only indirect evidences to support this
fact that DNA is the genetic material. The indirect evidences
are:
1. Localization 4. Effect of mutagens
2. Amount of DNA 5. Composition
3. Stability 6. Integrity

1. LOCALISATION
All cells of the living organisms have DNA. Particularly
located in the chromosomes fuelgen reaction is an evidence to
show that major component of chromosome is DNA which
performs the duty of the genetic material.
 2. AMOUNT OF DNA
The quality of DNA depends on the number of
chromosomes and the sets of chromosomes. Sex cells have
haploid number of chromosomes. Male and female sex cells
fuse together and become a zygote with diploid number of
chromosomes. In polyploidy cells the amount of DNA increases
with the increase in the number of sets of chromosomes. The
effect of extra chromosomes such as Klinefelter’s syndrome and
21-trisomy also prove that DNA is the genetic material

3. STABILITY
Studies on chromosomes reveal that genes are arranged in
a linear manner in all the chromosomes. Genes are nothing but
the series of polynucleotide. Genes are stable compounds.
Genes- DNA replicate without committing any mistakes, in each
and every cell division. Even then it is stable; rarely does it
change causing mutations.
4. EFFECT OF MUTAGENS
Mutations may occur spontaneously or through the effect
of mutagens such as radiation, chemicals etc. mutation alters
the chemical structure of DNA. Mutation is inherited by the
offspring which show phenotypic variations. These effects of
mutagens also give evidence to the fact that DNA is the genetic
material.

5. COMPOSITION
Structure of DNA is the same in all the organisms and
the other higher organisms. Composition of DNA is similar in
organisms of the same species and they inter breed. It is
different in organisms belonging to different species and they do
not interbreed.
6. INTEGRITY
DNA maintains integrity during cell metabolism and
also during embryonic development of the organisms. But
due to crossing over variations have been produced within
the species. Mutation disrupts the proper functioning of the
DNA.
The direct evidences of microorganisms and indirect
evidences of other higher organisms prove that DNA as the
stable genetic material which will survive in this biosphere
with all sorts of environmental hazards.

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