A

SEMINAR

ON

EVIDENCES OF NUCLEIC ACID

AS

“GENETIC MATERIAL”

SUBMITTED BY

PRIYANKA GUPTA

M.Sc MICROBIOLOGY (II SEMESTER)

SESSION 2018-2019

DEPARTMENT OF MICROBIOLOGY

KANYA GURUKUL CAMPUS

JWALAPUR (HARIDWAR)

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Contents

1. Genetic material 3

2. Properties of Genetic Material 3

3. Evidence from Bacteria 4-5

4. Evidence from Bacteriophage 6-7

5. Evidence from Conjugation 7-8

6. Evidence from RNA Viruses 8-9

7. Reference 10

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Genetic material
 These are those material of a cell or organism that are found in
the nucleus , mitochondria and cytoplasm that play a significant
role in determining the structure and nature of cell substance and
are capable of self-replication and variation.
 By the early 1900’s ,many biochemists had performed a variety of
experiments to find out whether protein or nucleic acid was the
genetic material.
 Some thought that proteins might be the genetic material as they
were abundant , diverse and complex type of molecules.
 But many experiments demonstrated that DNA instead of RNA
and protein is the genetic material in most microorganism and
RNA was found to be the genetic material in some viruses.

Properties of Genetic material

 Must carry information for cracking the genetic code .
 Must able to self- replicate for DNA replication .
 Must allow for information to change and capable of storing
hereditary information .
 Must be involved in gene - function to govern the expression of
phenotype .

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1. Evidence from Bacteria –
(a) Frederick Griffith : “ Bacterial Transformation’’
 In 1928 , British Bacteriologist Frederick Griffith gave an experimental
evidence that the DNA was the genetic material .
 He conducted a series of experiments using Streptococcus pneumoniae
and mice .
 In these experiment, he actually wasn’t trying to identify the genetic
material , but rather trying to develop a vaccine against pneumonia .
 He took 2 types of a bacterial strain , that causes pneumonia in humans
and other animals .

(i) R- strain :- It forms rough colonies when grown in petri – dish and lacks
LPS capsule i.e..they are non – virulent means they did not cause any disease
when injected into a mouse .

(ii) S- strain :-It forms smooth colonies when grown in petri – dish and possess
a LPS capsule that protects it from host immune system i.e..they are virulent
means they cause disease when injected into a mice .

Serotype Morphology of colony Capsule Virulence

Rll Rough Absent Non- virulent

SIII Smooth Present Virulent

He injected these strains into a mice in following 4 combinations and recorded

the results .

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 Result : From the 4th step , Griffith concluded that R – strain bacteria must
have taken up from the heat – killed S bacteria and called it a “transforming
principle’’ which allowed R- strain to transform into S –strain and become
virulent .

(b) Identification of transforming principle –

 As Griffith could not identify the nature of transforming substance , So in

1944 , three Canadian and American researchers –O. Avery , M.
McCarty and C. Macleod set out an experiment to identify which
organic molecule are causing the “transformation”.

 He separate the cell constituents in extract of virulent SIII using different

enzymes such as DNAse , RNAse , protease that hydrolysed DNA ,RNA
and proteins and then add or mixed this extract with RII-strain and
injected into mouse to see the transformation .

S.No Extract applied in RII Culture Transformation

1. DNA extract from heat killed SIII. YES

2. DNA extract from heat killed SIII treated with NO

DNAse enzyme which hydrolyse DNA.

3. DNA extract from heat killed SIII treated with YES

RNAse enzyme which hydrolse RNA .

4. DNA extract from heat killed SIII treated with YES

protease enzyme which hydrolyse proteins .

Result -: From their experiment ,they concluded that DNA is the genetic
material not the protein .

Because , the transformation occurred only when DNA was present in the
extract as in step 1,3 and 4 whereas no transformation occurred in step 2 due to
DNAse enzyme that hydrolyse DNA.

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2. Evidence from Bacteriophage -:
 In 1952, Alfred Hershey and Martha Chase carried out several
experiments on bacteriophage T2 that proved DNA to be the genetic
material of T2, which used E.coli as its host .

 T2 bacteriophage are composed of head coat and tail made up of protein

and DNA was found inside the protein coat .
 They showed that during reproduction of phages,the DNA of the phages
entered the host cell whereas the proteinaceous head and tail portion left
outside the host cell.
 Hence, it is strongly implied that the genetic information necessary for
viral reproduction was present in DNA.
 For this experiment , they prepared 2 culture medium for the growth of
E.coli that contained phosphorus and sulphur .

In Culture Ist -: Bacteriophage was grown in medium containing isotopic

phosphorus (32P) to make DNA radioactive.

In Culture IInd -:Bacteriophage was grown in medium containing isotopic

sulphur (35S) to make protein radioactive .

 Then poured the cell suspension of E.coli onto the growth medium.
 After bacterial growth,32P and 35S were incorporated into the bacterial
cell and phages were allowed to infect bacteria.
 After infection had taken place,each culture was subjected in a blender and
centrifuged to separate the phage from the bacteria.
 Then radioactivity of the phage and cell was measured.

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 Fig : Genetic material is transferred by DNA only

Result-:
DNA is the genetic material not protein.

3.Evidence from Bacterial Conjugation-:

 Conjugation is one of the method of transfer of DNA from donor
bacterium to the recipient bacterium through conjugation-tube that forms
between them.
 It was discovered by Lederberg and Tatum .
 The donor cell also called male cell (F+), contains fertility factor whereas
recipient cell also called female cell (F-) that lacks fertility factor.
 The F-factor is a mini circular DNA molecule and a segment of plasmid
DNA.

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  In 1946 ,Lederberg and Tatum demonstrated that when (F+) strain of
E.coli was mixed with (F-) strain ,then the (F-) cells were converted to
(F+).

(F+) + (F-) ( F+ )cells

Result -:
(F+) / (F-) (F+) / (F+)
From this experiment it was proved that DNA is the genetic material.

4. Evidence from RNA Viruses -:

The genome of viruses may be DNA and RNA .
Most of the plant viruses have RNA as their genetic material .
Fraenkel – Conrat in 1957 conducted experiments on Tobacco Mosaic
Virus (TMV) to demonstrate that in some viruses RNA acts as genetic
material.
TMV is a small rod- shaped virus composed of a single molecule of RNA
enclosed in protein coat .

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They took 2 different strains of TMV (A,B) and then separate their RNA
from protein coat .

Type A strain – contain RNA of TMV (A) and protein of TMV (B) .
Type B strain_- contain RNA of TMV (B) and protein of TMV (A) .
 They reconstituted hybrid viruses by mixing the protein of one strain
with the RNA of the second strain and vice – versa .
 When the hybrid or reconstituted viruses were rubbed onto tobacco
leaves , the progeny viruses produced were always found to be of
RNA type .
 Thus , the genetic information of TMV is stored in the RNA not in the
protein .

Fig: Demonstration of RNA as genetic material in TMV

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Reference

 Dubey. R.C, and Maheshwari .D.K (2010), Textbook of

Microbiology, S.Chand and Company Limited,Ram Nagar, New
Delhi-55.Chapter 5, page no-157-159.
 www.slideshare.net
 www.biologydiscussion.com
 www.khanacademy.org
 www.scribd.com

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