DNA and RNA as Genetic Material
D
N
A
& RNA
As
G
enetic
Material
21
/
08
/
2020
1
B
Y
D
r
.
K
.
A
r
u
n
a
A
s
s
i
s
t
a
n
t
P
r
o
f
e
s
s
o
r
D
e
p
a
r
t
m
e
n
t
o
f
M
i
c
r
o
b
i
o
l
o
g
y
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.
The
biochemical
studies reveal
that
chromosomes
are
composed
of
proteins
(60%
)
and
DNA
(40%
)
21/08/2020
2
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
21/08/2020
3
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
p
r
e
s
en
t
i
n
l
a
r
ger
q
uantit
y
,
e
n
cod
e
m
or
e
an
d
v
ar
iet
y
of
information.
D
NA
i
s
p
o
l
y
me
r
o
f
o
n
l
y
4
d
i
ff
er
en
t
deoxy
r
ibon
u
c
leotides
(ATP,
CTP,
GTP
&
TTP)
and
is
present
in
smaller
quantity
Most
geneticists focused
on
“transmission
genetics”
and
passively
accepted
proteins
as
the
genetic
material
Is
the
Genetic
Material
Protein
or
DNA
4
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
There
are
some
direct
evidences
and
some
indirect
evidences
which
prove
DNA
as
Genetic
Material
D
i
r
e
c
t
e
v
i
d
e
n
c
e
s
c
o
m
e
f
r
o
m
:
•
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
21/08/2020
5
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
21/08/2020
6
06/19/13
7
21/08/2020
STEPS
IN
THE
EXPERIMENT
1
L
I
V
E
2
SIII
LIVE
3
H
K
S
III
RII
4
H
K
S
III
&
LIVE
RII
S
t
r
a
i
n
s
o
f
D
i
p
l
o
c
o
c
c
u
s
p
n
e
u
m
o
n
i
a
e
i
n
j
e
c
t
e
d
t
8
o
m
i
c
e
Griffith’s
E
x
per
i
m
e
n
t
06
/
1
9
/
13
9
RII
SIII
transformation
takes
place
in
step
4
give
clue
for
DNA
as
genetic
material
1
2
3
4
√
S
N
I
II
I
II
STEPS
Mouse
injected with
SIII
strain
Mouse
injected with
RII
strain
Mouse
injected with
Heat
Killed
SIII
strain
RESULT
Mouse
died
Mouse
survived
Mouse
survived
I
V
living
R
II
strain
Mouse
injected with
Heat
Killed
SIII
&
Mouse
died
&
from
V
i
t
s
b
l
o
o
d
l
i
v
e
S
I
I
I
√
strain
bacteria
recovered
Mouse
survived
V
I
Mouse
injected with
Heat
Killed
SIII+
living
RII
strain
+
DNase
enzyme
Mouse
injected with
Heat
Killed
SIII+
living
RII
strain
+
Protease
enzyme
M
o
u
s
e
d
i
e
d
o
f
√
P
n
eu
m
o
n
ia
S
T
E
P
S
A
N
D
R
E
S
U
L
T
S
O
F
G
R
I
F
F
I
T
H
’
S
EXPERIMENT
21/08/2020
1
0
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
21/08/2020
11
Avery,
MacLeod,
&
McCarty,
1944
repeated
Griffith’s
using
purified
cell
extracts
and
experiment of
transformation
concluded
Removal
of
all
protein
from
the
transforming
material
did
not
destroy
its
ability
to
transform
R
strain
cells
DNA-digesting
enzymes
destroyed
all
transforming
ability
The
transforming
material
is
DNA
21/08/2020
12
A
V
E
R
Y,M
ACLEOD AND MCCART
Y
(1944)
21/08/2020
13
Avery,
MacLeod
AND
McCarty
EXPERIMENT
21/08/2020
14
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
Avery et
al
(1944)
revealed
the chemical
nature
of
the
transforming
substance
to
be
DNA.
With
the
help of
experiment
they
showed
that
DNA
isolated
from
SIII
strain Bacteria could
confer
the
pathogenic
properties
to
R
II
strain
Bacteria
.
Two
conclusion
were
derived
1.Active
factor
is
DNA
which
can
cause
transformation
2.SIII
strain
contains the
Active
factor
21/08/2020
15
Hershey
AND
Chase
Experiment
21/08/2020
16
A
l
f
r
e
d
H
e
r
s
h
e
y
a
n
d
M
a
r
t
h
a
C
h
a
s
e
,
1
9
5
2
i
n
v
e
s
t
i
g
a
t
e
d
b
a
c
t
e
r
i
o
p
h
a
g
e
s
:
v
i
r
u
s
e
s
t
h
a
t
i
n
f
e
c
t
b
a
c
t
e
r
i
a
-
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
Life
Cycle
of
T-
2
Phage
Phage
is
made
of
DNA
and
protein
coat
Only
DNA
enters
in
the
Bacterial
cell
and
protein
coat
is
left
out
side
21/08/2020
17
EVENTS
WHICH
TAKE
PLACE
IN
LIFE
CYCLE
OF
BACTERIOPHAGE
Attachment
of
Bacteriophage
to
receptor
site
on
bacterial
cell
wall
by
its
tail
Bacterial
cell
wall
dissolves
at
the
point
of
contact
Phage
DNA
enters
into
Bacterial cell,
protein
coat
remains
outside
Phage
DNA
replicates
forming
many
copies
Assembly
of
phage
DNA
into
new
protein
shells
Lysis
or
breakdown
of
host cell
and
release
of
infective
phage
particle.
21/08/2020
18
H
E
R
S
H
E
Y
&
C
H
A
S
E
(
1
9
52
)
E
X
P
E
R
I
M
E
N
T
W
I
T
H
T
2
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
(
Only
in
Ghosts)
Bacteriophage
progeny
carried
only
radioactive
phosphorus
and
not
radioactive
sulphur
21/08/2020
19
21/08/2020
20
21/08/2020
21
HERSHAY
&
CHASE
CONCLUSION
As
ghosts
or
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
21/08/2020
22
-
21/08/2020
23
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
SOME
ORGANISMS
HAVE
RNA
AS
GENETIC
MATERIAL
Some
Viruses
have
RNA
as Genetic
Material
RNA
Virus
(
Ribo
Virus )
may
have
ssRNA
or
dsRNA
enveloped
by
Protein
Coat
Notable
human
diseases caused
by
RNA
viruses
include
SARS,
Influenza
,Hepatitis
C,
Measels,
Mumps
and
Rabies
Rous
Sarcoma
Virus
(causing
cancer
)
and
HIV
(causing
AIDS)
both
are
Retrovirus using
Reverse
Transcription
for
DNA
synthesis
in
host
cell
21/08/2020
24
EXAMPLES
OF
PLANT
RNA
VIRUSES
Tobbaco
Mosaic
Virus
(TMV
)
was
crystallized
by
Stanley
(1935)
for the
first
time
TMV
causes Tobacco
mosaic
disease
TMV can
also cause disease in
Tomato,
Pepper,
Petunia, Snapdragon,
Delphinium,
and
Marigold.
Papaya
Ring
Spot virus
(PRV)
Potato Leaf Roll Virus
(PLRV)
Holmesrib-grass Virus (HRV)
–Plantago
Alfa-alfa Mosaic Virus (AMV)
21/08/2020
25
T
M
V
HIV
HEPATITES
C
VIRUS
RNA
VIRUSES
21/08/2020
26
EVIDENCE
IN
FAVOUR
OF
RNA
AS
GENETIC
MATERIAL
A
Gierer
and
G
Schramm
(
1956
)
when
inoculated
to
ba
c
c
o
p
l
a
n
t
s
w
it
h
P
urif
i
e
d
RN
A
i
s
o
la
t
e
d
fr
o
m
T
M
V
-
Leisons
appeared on
leaves
of
healthy
plant
H
Fraenkel
Conrat
and
B
Singer
(1957)
separated
RNA
from
protein
of
TMV
in
first
step
.
In
second
step
reconstituted
virus
with
protein
from
mutant
strain
of
TMV
21/08/2020
27
Inoculated
Hybrid
TMV
into
healthy
tobacco
plant
Tobacco
Mosaic disease
appeared
T
M
V
p
r
o
ge
n
y
is
o
l
a
ted
f
r
o
m
d
isea
s
e
d
p
lan
t
s
h
o
w
e
d
parental
RNA
only
but
not
parental
proteins
This
provided
evidence
that
RNA
carries
information
for
assembly
of
Virus
particles
and
not
proteins
RNA
thus
is
the
Genetic
Material
in
TMV
21/08/2020
3
1
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
21/08/2020
29
Expression
is controlled
through
Central
Dogma
of
Molecular
Biology
involving Replication,Transcription
and
Translation
Results
of
Experiments
performed
by
Griffith’s
(1928),
Avery
et
al
(1944)
and
Hershey
and
Chase
(1952)
provide
direct
evidences
in
favour
of
DNA
as
Genetic
Material
There
are
some
indirect
evidences
also
in
favour
of
DNA
as Genetic
Material
In
some
organisms
RNA
is
the
Genetic
Material
Evidences
are
given
by
Experiments
on
TMV
21/08/2020
30
D
N
A
p
r
e
R
N
A
R
N
A
P
r
o
t
e
i
n
D
N
A
R
e
p
l
i
c
a
t
i
o
n
&
R
e
p
a
i
r
T
r
a
n
s
c
r
i
p
t
i
o
n
P
r
o
c
e
s
s
i
n
g
T
r
a
n
s
l
a
t
i
o
n
F
o
l
d
i
n
g
A
s
s
e
m
b
l
y
P
r
o
c
e
s
s
i
n
g
C
e
n
t
r
a
l
D
o
g
m
a
THE
MODERN
FRAME
WORK
OF
MOLECULAR
BIOLOGY
21/08/2020
31
21/08/2020
32
Genetic material plays a crucial role in the transmission of traits from one generation to the next. This article explores the significance of DNA and RNA as genetic material, highlighting key experiments and discoveries that demonstrate DNA's role in carrying hereditary information. From Mendel's hybridization experiments to more recent studies, the evidence overwhelmingly supports DNA as the primary carrier of genetic information.
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Presentation Transcript
DNA & RNA As Genetic Material BY Dr. K. Aruna Assistant Professor Department of Microbiology 21/08/2020 1
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 sometraits 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. The biochemical studies reveal that chromosomes are composed of proteins (60% ) and DNA (40% ) 21/08/2020 2
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 21/08/2020 3
Is the Genetic Material Protein or DNA 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 Most geneticists focused on transmission genetics and passively accepted proteins as the genetic material 4
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 There are some direct evidences and some indirect evidences which prove DNA as Genetic Material Direct evidences come from : Frederick Griffith s (1928) experiment on Bacterial Transformation Oswald Avery, Colin Macleod and MaclynMcCarty s (1944 ) experiment on Transformation Alfred D.Hershey and Martha Chase (1952) experiment on T- Even (2,4 ) Bacteriophage 21/08/2020 5
Frederick Griffith (1928) Studied Diplococcus pneumoniae, having Twostrains 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 21/08/2020 6
06/19/13 7
STEPS IN THEEXPERIMENT 1 LIVE SIII Strains of Diplococcus pneumoniae injected t8omice 2 LIVE RII 3 HK S III 4 H K S III & LIVERII 21/08/2020
Griffiths Experiment 1 RII SIII transformation takes place instep4 give clue for DNAas geneticmaterial 2 3 4 06/19/13 9
STEPS AND RESULTS OF GRIFFITHS EXPERIMENT SN I II III STEPS RESULT Mouse died Mouse survived Mousesurvived Mouse injected with SIII strain Mouse injected with RII strain Mouse injected with Heat Killed SIII strain Mouse injected with Heat Killed SIII & Mouse died & from IV living R IIstrain its blood live SIII strain bacteria recovered Mousesurvived Mouse injected with Heat Killed SIII+ living RII strain + DNaseenzyme Mouse injected with Heat Killed SIII+ living RII strain + Proteaseenzyme V VI Mouse died of Pneumonia 10 21/08/2020
CONCLUSION GRIFFITH S 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 21/08/2020 11
Avery, experiment of transformation MacLeod, & McCarty, 1944 repeated Griffith s using purified cell extracts and concluded Removal of all protein from the transforming material did not destroy its ability to transform R strain cells DNA-digesting enzymes destroyed all transforming ability The transforming material is DNA 21/08/2020 12
AVERY,M ACLEOD AND MCCART Y (1944) 21/08/2020 13
Avery, MacLeod ANDMcCarty EXPERIMENT Is based ontransformation 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 Proteaseand RNase transformation was recorded In case of DNase no transformation was recorded 21/08/2020 14
Avery et al (1944) revealed the chemical nature of the transforming substance to be DNA. With the help of experiment they showed that DNA isolated from SIII strain Bacteria could confer the pathogenic properties to R II strain Bacteria . Two conclusion werederived 1.Active factor is DNA which can cause transformation 2.SIII strain contains the Active factor 21/08/2020 15
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 21/08/2020 16
Life Cycle of T- 2 Phage Phage is made of DNA and proteincoat Only DNA enters in the Bacterial protein coat is left out side cell and 21/08/2020 17
EVENTS WHICH TAKE PLACE IN LIFE CYCLE OFBACTERIOPHAGE Attachment of Bacteriophage to receptor siteon bacterial cell wall by its tail Bacterial cell wall dissolves at the point ofcontact Phage DNA enters into Bacterial cell, protein coat remains outside Phage DNA replicates forming manycopies Assembly of phage DNA into new proteinshells Lysis or breakdown of host cell and release of infective phage particle. 21/08/2020 18
HERSHEY & CHASE (1952) EXPERIMENT WITH T2 BACTERIOPHAGE In culture I Bacteriophage was grown in medium containing Radioactive Phosphorus (32P))To make DNARadioactive In culture II Bacteriophage was grown in medium containing radioactive Sulphur (35S) 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 ( Only in Ghosts) Bacteriophage progeny carried only radioactive phosphorus and not radioactive sulphur 21/08/2020 19
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HERSHAY & CHASECONCLUSION As ghosts or 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 notproteins 21/08/2020 22
Summary of Hershey & Chase (1952 )experiment Bacteriophage DNA was labeled with radioactive phosphorus (32P) - - Bacteriophage protein was labeled with radioactive sulphur (35S) - Radioactive molecules weretracked - only the bacteriophage DNA (as indicated by the32P) entered the bacteria and was used to produce more bacteriophage Conclusion: DNA is the genetic material 21/08/2020 23
SOMEORGANISMS HAVE RNA AS GENETIC MATERIAL Some Viruses have RNA as GeneticMaterial RNA Virus ( Ribo Virus ) may have ssRNA or dsRNA enveloped by Protein Coat Notable human diseases caused by RNA viruses include SARS, Influenza ,Hepatitis C, Measels, Mumps and Rabies Rous Sarcoma Virus (causing cancer ) and HIV (causing AIDS) both are Retrovirus using Reverse Transcription for DNA synthesis in host cell 21/08/2020 24
EXAMPLES OF PLANT RNA VIRUSES Tobbaco Mosaic Virus (TMV) was crystallized by Stanley (1935) for the firsttime TMV causes Tobacco mosaic disease TMV can also cause disease in Tomato, Pepper, Petunia, Snapdragon, Delphinium, and Marigold. Papaya Ring Spot virus (PRV) Potato Leaf Roll Virus (PLRV) Holmesrib-grass Virus (HRV) Plantago Alfa-alfa Mosaic Virus (AMV) 21/08/2020 25
RNAVIRUSES HEPATITES C VIRUS TMV HIV 21/08/2020 26
EVIDENCE IN FAVOUR OF RNA AS GENETIC MATERIAL A Gierer and G Schramm ( 1956 ) when inoculated tobacco plants with Purified RNA isolated from TMV - Leisons appeared on leaves of healthy plant H Fraenkel Conrat and B Singer (1957) separated RNA from proteinof TMV in first step. In second step reconstituted virus with protein from mutant strain ofTMV 21/08/2020 27
Inoculated Hybrid TMV into healthy tobacco plant Tobacco Mosaic disease appeared TMV progeny isolated from diseased plant showed parental RNA only but not parentalproteins This provided evidence that RNA carries information for assembly of Virus particles and not proteins RNA thus is theGenetic Material in TMV 31 21/08/2020
SUMMARY OF THE PRESENTAION ON DNA AS GENETICMATERIAL 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 undergomutation 21/08/2020 29
Expression is controlled through Central Dogma of Molecular Biology involving Replication,Transcription andTranslation Results of Experiments performed by Griffith s (1928), Avery et al (1944) and Hershey and Chase (1952) provide direct evidences in favour of DNAas Genetic Material There are some indirect evidences also in favour of DNAas Genetic Material In some organisms RNA is the Genetic Material Evidences are given by Experiments on TMV 21/08/2020 30
THE MODERN FRAME WORK OF MOLECULAR BIOLOGY Processing Transcription Translation preRNA DNA RNA Replication & Repair Protein Folding Assembly Processing DNA Central Dogma 21/08/2020 31
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