The document discusses evidence that DNA is the genetic material:
1) Griffith's experiment showed bacterial transformation from non-virulent to virulent strains upon uptake of DNA from heat-killed bacteria.
2) Avery, MacLeod and McCarty's experiment showed transformation only occurred when DNA was present, as DNase treatment prevented transformation.
3) Hershey and Chase's experiment using radioactive labels showed that only bacteriophage DNA, not protein, entered bacteria to produce new phage particles.
2. • It is well known fact that transmission of traits takes place
from one generation to other. The offsprings are similar to
both the parents in some traits
• Gregor Johann Mendel (1866 ) on the basis of his
hybridization experiments on Sweet pea gave the idea that
transmission of traits over generations take place through
Factor or Determiner or Gene which carries information for
expression of trait or phenotype.
• Genes are present on the chromosomes which are distributed
equally into the two daughter cells during cell division.
11/11/162
3. Genetic material must be capable of
• Replication ( Make its copy )
• Storage of information for expression of trait
• Control expression of traits
• Change in controlled way (undergo mutation)
• Must be stable
11/11/163
4. • Topic of Discussion till 1944 was which chromosomal
component DNA or Protein carries hereditary
information or is the genetic material
• Untill 1940 Proteins were considered as genetic
material as Proteins are polymer of 20 protein amino
acids and present in larger quantity, encode more and
variety of information.
• DNA is polymer of only 4 different
deoxyribonucleotides (ATP, CTP, GTP & TTP) and is
present in smaller quantity
11/11/164
Is the Genetic Material Protein or DNA
5. But On the basis of certain experiments conducted from time to
time ,it was ultimately demonstrated that DNA carries genetic
information and not the proteins
11/11/165
Direct evidences come from :
• Frederick Griffith’s (1928) experiment on Bacterial
Transformation
• Oswald Avery, Colin Macleod and Maclyn McCarty’s
(1944 ) experiment on Transformation
• Alfred D. Hershey and Martha Chase (1952 )
experiment on T- Even (2,4 ) Bacteriophage
6. Frederick Griffith (1928) Studied Diplococcus
pneumoniae, having Two strains
• SIII strain was virulent, possessed a lipopolysaccharide
capsule and could kill mice by causing disease Pneumonia and
made round colonies on a culture plate
• RII strain was avirulent and lacked a Lipopolysaccharide (LPS)
capsule, growing in rough–shaped colonies on a culture plate
11/11/166
8. S N STEPS RESULT
I Mouse injected with SIII strain Mouse died √
II Mouse injected with RII strain Mouse survived
III Mouse injected with Heat Killed SIII
strain
Mouse survived
IV Mouse injected with Heat Killed SIII &
living R II strain
Mouse died & from
its blood live SIII √
strain bacteria
recovered
V Mouse injected with Heat Killed SIII+
living RII strain + DNase enzyme
Mouse survived
VI Mouse injected with Heat Killed SIII+
living RII strain + Protease enzyme
Mouse died of √
Pneumonia
STEPS AND RESULTS OF GRIFFITH’S
EXPERIMENT
11/11/168
9. GRIFFITH’S CONCLUSION
•On the basis of Result of step IV Griffith concluded that there was
transformation of Avirulent RII type to Virulent SIII type by picking up
the genetic material encoding the LPS capsule from the Heat Killed S
III .
•This bacterial transformation clearly shows the role of DNA as Genetic
Material and is further confirmed by results of step V which shows no
transformation as DNase digests DNA and step VI again shows
transformation as protease only digests protein
• DNA-digesting enzymes destroyed all transforming ability
•The transforming material is DNA
11/11/169
10. 11/11/1610
Avery, MacLeod AND McCarty
EXPERIMENT
Is based on transformation
Cell free extract of SIII strain Bacterium was subjected to
DNase,RNase and Protease
Each treated extract was mixed with RII and mixture
injected to mouse to see transformation.
In case of Protease and RNase transformation was recorded
In case of DNase no transformation was recorded
11. 11/11/1611
Hershey AND Chase Experiment
•Alfred Hershey and Martha Chase, 1952 investigated
bacteriophages: viruses that infect bacteria
•- the bacteriophage was composed of only DNA and
protein
•- they wanted to determine which of these molecules is the
genetic material that enters into the bacteria
12. HERSHEY & CHASE (1952) EXPERIMENT WITH T2
BACTERIOPHAGE
• In culture I Bacteriophage was grown in medium containing Radioactive
Phosphorus (32
P))To make DNA Radioactive
• In culture II Bacteriophage was grown in medium containing radioactive
Sulphur (35
S) To make proteins Radioactive
• Both kinds of Bacteriophage particles were allowed to infect Bacteria
• The infected bacteria were observed for radioactivity
• Radioactive Phosphorus was found with bacterial cells
• Radioactive Sulphur was not traced in bacterial cells
• Bacteriophage progeny carried only radioactive phosphorus and not
radioactive sulphur
11/11/1612
14. HERSHAY & CHASE CONCLUSION
• As coat of Bacteriophage were not labelled with 32P and
only with 35S.
• The results of experiment clearly indicate that only DNA
and not the proteins enter the bacterial cell. Protein coat
is left outside . The DNA entering the host cell carries all
the genetic information for synthesis of new phage
particle.
• This certainly proves that DNA is the genetic material in
Bacteriophage and not proteins 11/11/1614
15. 11/11/1615
- Bacteriophage DNA was labeled with radioactive
phosphorus (32
P)
- Bacteriophage protein was labeled with radioactive
sulphur (35
S)
- Radioactive molecules were tracked
- only the bacteriophage DNA (as indicated by the 32
P)
entered the bacteria and was used to produce more
bacteriophage
Conclusion: DNA is the genetic material
Summary of Hershey & Chase (1952 ) experiment
16. INDIRECT EVIDENCES FOR DNA AS
GENETIC MATERIAL
• Presence of DNA regularly in nuclei of all types of cells
• Equal amount of DNA is present in all cells of an organism
• Amount of DNA is proportional to the ploidy of cell . Haploid
cells have ½ the amount of DNA than diploid cell.
• Nuclear Division occurs only after DNA duplication during
S phase of Interphase
• Variation in Diploid amount of DNA amongst different
species
11/11/1616
17. SUMMARY OF THE PRESENTAION ON
DNA AS GENETIC MATERIAL
• Gene (Mendelian Factor) carries information for
expression of Phenotype.
• Genes are present on chromosomes.
• The chromosomes contain 60% protein and 40% DNA.
• Genetic Material must be stable, capable of replication,
storing information for expression and undergo mutation.
• Expression is controlled through Central Dogma of
Molecular Biology involving Replication ,Transcription and
Translation.
11/11/1617
18. DNA preRNA RNA
Protein
DNA
Replication
& Repair
Transcription Processing Translation
Folding
Assembly
Processing
Central Dogma
THE MODERN FRAME WORK OF
MOLECULAR BIOLOGY
11/11/1618