Unit 9 Genetic Material

UNIT 9
GENETIC MATERIAL

Structure
9.1 Introduction 9.5 Transformation
Objectives Experiments for
9.2 Types of Genetic Material Confirmation of DNA as
9.3 Deoxyribonucleic Acid Genetic Material
(DNA) Griffith’s Experiment

Discovery of DNA Avery’s Experiment

Structure of DNA Hershey-Chase Bacteriophage

9.4 Ribonucleic Acid (RNA) Experiment

9.6 Summary
9.7 Terminal Questions
9.8 Answers

9.1 INTRODUCTION
This is the first unit of second volume. Here we will study about the structure of
genetic material which is responsible for various activities of living organisms
and passing of characters from one generation to other. Cells use different
types of molecules to store diverse types of information to keep themselves
alive.

Most of you are familiar with nucleic acids which are molecules of great
importance to cell because of their role in storage, transmission and
expression of genetic information. The two major nucleic acids i.e. DNA
(deoxyribonucleic acid) and RNA (ribonucleic acid) play an important role in
storage of genetic information and expression of the information via protein
synthesis. Both DNA and RNA differ in their chemical nature and functions
they perform in the cell. In most of the cells, DNA is present within the nucleus.
DNA acts as the genetic material in majority of living organisms. In some
organisms such as viruses, RNA is the main hereditary material. In some
organisms such as bacteriophages, both DNA as well RNA carry genetic
information for the cells.

In this unit we are going to discuss the genetic material of the cell because it
carries and stores fundamental information necessary for life of every living 9
Block 3 DNA-Blueprint of Life
organism. You will be studying about structure of DNA and RNA, their roles in
organism and some experiments that provided the evidence that DNA is the
hereditary material present in plants. This is an introductory unit for DNA
structure.

Objectives
Objectives
After studying this unit, you would be able to:

describe the types of genetic material found in living organisms;

describe the structure of DNA and RNA;

provide evidence about DNA as genetic material through transformation

experiments; and

appreciate a detailed account of classical experiments by Griffith, Avery

and Hershey-Chase.

9.2 TYPES OF GENETIC MATERIAL

Cells contain material or matter that controls inheritance of traits from one
generation to the next. It is also able to convey its effect through the formation
and functioning of the traits. This material is referred as genetic material. In
genetic material, the hereditary information is present in coded form in genes.
Thus the genetic material of an individual consists of several thousands of
genes. The genetic material possess certain properties such as stability,
expressed when required, replicates and is capable of being transmitted from
parent to progeny without change. The replicated genetic material must be
transferred from a cell to its daughter cells and from one generation to the
next. In most of the living organisms, the genetic information is stored in the
nucleotide sequences of DNA molecules. Both DNA and RNA carry the
genetic information in living organisms. In most of the organisms, DNA is the
genetic material, however in some viruses, bacteriophages, RNA has been
found as the genetic material. It has been noted that in organisms where DNA
is absent, RNA functions as genetic material.

Structure of Nucleic Acids (Polynucleotides)

Nucleic acid molecule is a long chain polymer (polynucleotide) that is

composed of monomeric units, called nucleotides. These monomer units of
nucleotide are joined by the formation of phosphodiester bonds (a diester
bond is one that involves two ester bonds). These phosphodiester bonds are
formed between any two neighboring nucleotides.

Each nucleotide consists of three components- nitrogen-containing bases, a

five-carbon sugar, and a phosphate group. A phosphate group and nitrogen
containing aromatic base is attached to five carbon sugar. The nitrogenous
base is either a purine or a pyrimidine. The five-carbon sugar is either a ribose
(in RNA) or a deoxyribose (in DNA) molecule. The phosphate is joined by a
phosphoester bond to the 5ˊ carbon of the sugar. The base is attached at the
10 1ˊ carbon (Fig.9.1).
Unit 9 Genetic Material

Fig. 9.1: Polynucleotide chain showing phosphodiester bonds.

If we remove phosphate group from the nucleotide, remaining base-sugar is
called nucleoside. The purine or pyrimidine thus occurs as free base, the
nucleoside or nucleotide (Fig.9.2).

Fig. 9.2: A short-hand notation of a polynucleotide with four nucleotides (1ˊ to 5ˊ

represent numbers of five carbon atoms of pentose sugar). 11
Block 3 DNA-Blueprint of Life
The phosphate group is attached by a phosphodiester bond to the 5ˊ carbon of
one nucleotide becomes linked by a second phosphoester bond to the 3ˊ
carbon of the next nucleotide. The condensation of H and OH groups coming
from sugar and the phosphate results in linkage called phosphodiester bond.
The polynucleotide is formed by such interactions. It shows the presence of 3ˊ
hydroxyl group at one end and 5ˊ hydroxyl group at the other end.

9.3 DEOXYRIBONUCLEIC ACID (DNA)

As you know that DNA is the genetic material that gets inherited from parent to
offspring. Most DNA is located in nucleus of the cell (called nuclear DNA), but
a small amount of DNA can also be found in the mitochondria (called
mitochondrial DNA). DNA is organized structurally into chromosomes wound
around nucleosomes as part of those chromosomes. Functionally, it's
organized into genes that carry information for observable traits. Genes are
small sections of the long chain of DNA. Gene is considered as functional unit
of heredity. Genes possess instructions to make molecules called proteins
(Fig.9.3).

Fig 9.3: Double helix structure of DNA. Chemical structure for three base pair is
12 shown.
Unit 9 Genetic Material
DNA is made of 4 types of nucleotides which are arranged variously to form
long chain molecules. The DNA molecule consists of two strands that wind
around one another to form a shape known as a double helix. Each strand has
a backbone made of sugar (deoxyribose) and phosphate groups. To each
sugar is attached one of bases - adenine (A), cytosine (C), guanine (G), and
thymine (T). The two strands are held together by hydrogen bonds between
the bases; adenine (A) binds with thymine (T), and cytosine (C) binds with
guanine (G). Each base is attached to a sugar molecule and a phosphate
molecule. This pairing results in formation of base pairs. Base, sugar, and
phosphate together form a nucleotide. Nucleotides are arranged in such a way
that two long strands with their sugar –phosphate are on outside of the helix
and their bases are on the inside. This spiral is called a double helix. You can
imagine the structure of the double helix like a ladder, with base pairs running
through the middle like rings and sugar and phosphate molecules along the
outside. Helix is absolutely regular and it can be distinguished by a major and
minor groove.

9.3.1 Discovery of DNA

DNA was first identified in the late 1860s by Swiss chemist Friedrich Miescher.
Decades after Miescher's discovery, other scientists-Phoebus Levene, Erwin
Chargaff, Albrecht Kossel, Maurice Wilkins and Rosalind Franklin carried out a
series of researches that revealed additional details about the DNA molecule,
including its primary chemical components and structure. The scientific
foundation provided by these pioneers, provided preliminary information to
Watson and Crick and they proposed that DNA molecule exists in the form of
a three-dimensional double helix in 1953.

The apparent first observation of a substance later identified as nucleic acid

was made by Justus Liebig, who in 1847 reported the presence of an acidic
material in a filtrate obtained from beef muscle, which he then named
“inosinic acid” (from the Greek word inos, fiber, and more specifically those of
muscle). Twenty years later Gregor Mendel presented the results of his work
on inheritance in pea plant, Friedrich Miescher, became interested in
isolating and characterizing the chemical components present in the nucleus
of a cell. He chose leukocytes (white blood cells) isolated from the pus coated
surgical bandages. The extract obtained had chemical properties unlike any
previously discovered molecule, which had higher phosphorus content.
Miescher realized that he had discovered a new substance which he named
as “nuclein”. Finally, in 1889, Richard Altmann succeeded in removing
protein associated with nucleins and he named the remaining acidic material
as “nucleic acid” (Nucleinsäure). This scientific achievement opened the
gateway to investigate nucleic acids apart from their accompanying proteins.

Followed by Albrecht Kossel, Altmann’s used several methods to determine

the main chemical components of the nucleic acids. Kossel’s found that
hydrolysis of nucleic acids produced large quantities of phosphoric acid and
specific sets of other molecules depending on the original material used as the
source. Such compounds included purine -adenine, guanine, and 13
Block 3 DNA-Blueprint of Life
pyrimidine bases—thymine, uracil, and cytosine. By the end of the
century, a variety of nucleic acids with different elemental compositions were
known. Two main kinds of nucleic acids were distinguished i.e. the yeast
nucleic acid, believed to be typical of plants (also called “phytonucleic”), and
that obtained from the thymus gland (“thymonucleic”, or occasionally
“zoonucleic”), which was seen as particularly associated to animals. The
main differences detected between these two types of nucleic acids (they were
-RNA and DNA) were in composition regarding one of the pyrimidine bases
(thymine in thymonucleic versus uracil in yeast).

Phoebus Levene, a Russian physician in collaboration with his student J. A.

Mandel extracted DNA from the thymus. Later Kossel identified the base as
“thymine.” They concluded that linear complexes consist of one of a
phosphoric acid, a carbohydrate, and a base that bind in such a way that they
form a polyphosphoric acid. The base is probably bound to the sugar moiety in
glycosidic form”. Levene was the first to discover the order of the three major
components of a single nucleotide (phosphate-sugar-base), carbohydrate
component of DNA and RNA. Levene proposed that nucleic acids were
composed of a series of nucleotides, and that each nucleotide was in turn
composed of just one of four nitrogen-containing bases, a sugar molecule, and
a phosphate group. He also proposed a tetranucleotide structure, in which
the nucleotides were always linked in the same order (i.e., G-C-T-A-G-C-T-A
and so on).

In 1962 Watson, Crick, and Wilkins jointly received the Nobel Prize in
Physiology or Medicine for their determination of the structure of
deoxyribonucleic acid (DNA). In 1953 Rosalind Franklin and Maurice
Wilkins used X-ray diffraction technique called X-ray crystallography to figure
out the structure of DNA. X-rays were beamed through crystals of
the DNA molecule and photographic film was used to record the scattering of
X rays. The DNA crystals resulted in a cross shape on the X-ray film which is
typical of a molecule with a helix shape. They found that DNA molecules are
helical with two periodicities along their long axis, a primary one of 3.4 Å and a
secondary one of 34 Å.

Box 9.1: Unsung Hero Rosalind Elsie Franklin

nth
Rosalind Elsie Franklin (Fig. 9.4) was born in London, England on 16 of April 1920.
Rosalind Franklin was exceptionally bright and by the age of 15 she knew that she
wanted to be a scientist. Her father actively discouraged her interest since it was very
difficult for women to have such a career. However, with her excellent education
from St. Paul's Girls' School which taught physics and chemistry to girls. Franklin
entered Cambridge University in 1938 to study chemistry. When she graduated,
Franklin was awarded a research scholarship to do graduate work. She spent a year
in R.G.W. Norrish's lab without great success. Norrish recognized Franklin's potential
but he was not very encouraging or supportive towards his female student. When
offered the position as an assistant research officer at the British Coal Utilization
Research Association (CURA), Franklin gave up her fellowship and took the job.

CURA being a young organization, was less firm on the way research was to be done.
14 Franklin worked quite independently, a condition that suited her. Franklin worked for
Unit 9 Genetic Material
CURA until 1947 and published a number of papers on the physical structure of coal.
Then she moved to Paris for doing research work. An old friend introduced her to
Marcel Mathieu who directed most of the research in France. He was quite impressed
with Franklin's work and offered her a job as a "chercheur" in the Laboratoire Central
des Services Chimiques de l'Etat. There she learned X-ray diffraction techniques from
Jacques Mering.

In 1951, Franklin was presented with a 3-year research scholarship at King's College.
Franklin began working as a research associate in the biophysics unit, where director
John Randall used her expertise and X-ray diffraction techniques (mostly of proteins
and lipids in solution) on DNA fibers. Here Maurice Wilkins was already using X-ray
crystallography to try to solve the DNA problem at King's College. Studying DNA
structure with X-ray diffraction, Franklin and her student Raymond Gosling made an
amazing discovery: They took pictures of DNA and discovered that there were two
forms of it, a dry "A" form and a wet "B" form. One of their X-ray diffraction pictures of
the "B" form of DNA, known as Photograph 51, became famous as critical evidence in
identifying the structure of DNA. The photo was acquired through 100 hours of X-ray
exposure from a machine Franklin herself had refined. From this she deduced the
basic dimensions of DNA strands, and that the phosphates were on the outside of
what was probably a helical structure.

Fig 9.4: Rosalind Franklin (1920-1958).

John Desmond Bernal, United Kingdom’s one of the most well-known and
controversial scientists and a pioneer in X-ray crystallography, spoke highly of Franklin
around the time of her death in 1958. "As a scientist Miss Franklin was distinguished
by extreme clarity and perfection in everything she undertook," he said. "Her
photographs were among the most beautiful X-ray photographs of any substance ever
taken. Their excellence was the fruit of extreme care in preparation and mounting of
the specimens as well as in the taking of the photographs."
She presented her data at a lecture in King's College that James Watson was
attending. In his book The Double Helix, Watson admitted to not paying attention at
Franklin's talk and not being able to fully describe the lecture and the results to Francis
Crick. Watson and Crick were at the Cavendish Laboratory and had been working on
solving the DNA structure. Franklin did not know Watson and Crick as well as Wilkins
did and never truly collaborated with them. It was Wilkins who showed Watson and
Crick the X-ray data Franklin obtained. The data confirmed the 3-D structure that
Watson and Crick had theorized for DNA.

Despite her cautious and diligent work ethic, Franklin had a personality conflict with
colleague Maurice Wilkins, one that would end up costing her greatly. In January
1953, Wilkins changed the course of DNA history by disclosing without Franklin's
permission or knowledge her Photo 51 to competing scientist James Watson, who was
working on his own DNA model with Francis Crick at Cambridge.
15
Block 3 DNA-Blueprint of Life
Upon seeing the photograph, Watson said, "My jaw fell open and my pulse
began to race," according to author Brenda Maddox, who in 2002 wrote a book
about Franklin titled Rosalind Franklin: The Dark Lady of DNA.

The two scientists did, in fact, use what they saw in Photo 51 as the basis for their
famous model of DNA, which they published on March 7, 1953, and for which they
received a Nobel Prize in 1962. Crick and Watson were also able to take most of the
credit for the finding: When publishing their model in Nature magazine in April 1953,
they included a footnote acknowledging that they were "stimulated by a general
knowledge" of Franklin's and Wilkins' unpublished contribution, when in fact, much of
their work was rooted in Franklin's photo and findings. Randall and the Cambridge
laboratory director came to an agreement and both Wilkins' and Franklin's articles
were published second and third in the same issue of Nature. Still, it appeared that
their articles were merely supporting Crick and Watson’s model.
Franklin left Cambridge in 1953 and went to the Birkbeck lab to work on the structure
of tobacco mosaic virus. She published a number of papers on the subject and she
actually did a lot of the work while suffering from cancer. She died from cancer in
1958. In the fall of 1956, Franklin discovered that she had ovarian cancer. She
continued working throughout the following two years, despite having three operations
and experimental chemotherapy. She experienced a 10-month remission and worked
up until several weeks before her death on April 16, 1958, at the age of 37.

In 1962, the Nobel Prize in Physiology or Medicine was awarded to James Watson,
Francis Crick, and Maurice Wilkins for solving the structure of DNA. The Nobel
committee does not give posthumous prizes.

9.3.2 Structure of DNA

In 1953 J.D. Watson and F.H.C. Crick (Fig. 9.5) proposed a double helical
DNA was first
crystallized in the late structure of DNA. According to them the two polynucleotide chains are
70's -the 1953 X-ray connected by hydrogen bonds and running in opposite directions. In a DNA
data were from DNA double helix two polynucleotide chains are coiled about the same axis in such
fibers. So, the real
a manner that they can separate from one another only be uncoiling. Bases
"proof" for the
Watson-Crick model are present in a plane at right angle to the long axis.
of DNA came in 1982
after the B-form of
DNA was crystallized
and the X-ray pattern
was solved.

16 Fig 9.5: J.D. Watson and F.H.C. Crick

Unit 9 Genetic Material
DNA is a stable molecule. Two polynucleotide chains which run in opposite
directions have complementary base sequences.

You will realize that one most important feature to know that within the helix an
adenine is always adjacent to thymine in the other strand and similarly
guanine is always adjacent to cytosine in one polynucleotide. This means
adenine is linked to thymine and cytosine to guanine. The distance between
two base pairs is 3.4 Å and there are 10 base pairs in each turn. The molecule
has a diameter of 20 Å in the molecule. Purines which are adenine and
guanine need a little higher space as compared to two pyrimidines i.e.
cytosine and thymine. When a pyrimidine always pairs with a purine, space
could remain constant (Fig. 9.6). Normally they are written as A, G, C and T
(Fig.9.7).

Fig. 9.6: A double helix model of DNA as proposed by Watson and Crick.

Fig. 9.7: Chemical structure of purines and pyrimidines.

Erwin Chargaff, an Austrian biochemist, expanded Levene's work and found

additional information of the structure of DNA. Chargaff found that nucleotide
composition of DNA varies among species. He concluded that almost all DNA-
no matter what organism or tissue type it comes from but maintains certain 17
Block 3 DNA-Blueprint of Life
properties; even as its composition varies. Using paper chromatography, he
found that the amount of adenine (A) is usually similar to the amount of
thymine (T), and the amount of guanine (G) usually approximates the amount
of cytosine (C). In other words, the total amount of purines (A + G) equals the
total amount of pyrimidines (C + T) (known as "Chargaff's rule"). All DNA
follow Chargaff's Rule that states that the total number of purines in a DNA
molecule is equal to the total number of pyrimidines.

A+T=G+C

The analysis of DNA obtained from different organs of body of the same
individual or from different individuals belonging to the same species showed
no differences in the relative composition of different bases. The studies
suggested that:

• Number of adenine molecules is equal to number of thymine molecules.

• Number of cytosine molecules is equal to number of guanine molecules.

Chargaff’s rule may be represented as follow:

A=T: G=C; A+T= G+C

A+G=T+C
A+G/T+C=1; But
A+T ≠ G+C
i.e., A=T/G+C = constant

For example, this ratio is 0.4 for Bacillus whereas human DNA has the A:G
ratio of 1.56

SAQ 1
a) Fill in the blanks:

i) DNA consists of ………………. types of nucleotides.

ii) DNA is structurally organized into ………………….. .

iii) Justus Liebig isolated a substance from beef muscle and named it
as ……………….. .

iv) Miescher discovered a new substance and named it as

……………….. .

b) Write the full form of the following:

A, G (purines) and C, T (pyrimidines)

9.4 RIBONUCLEIC ACID (RNA)

Generally RNA is formed from DNA during transcription. It carries genetical
instructions that determine the sequence of amino acids in protein formed
during translation. RNA is found as single stranded molecule. RNA consists of
18 ribose nucleotides attached by phosphodiester bonds. The four bases present
Unit 9 Genetic Material
in RNA, are adenine, guanine, uracil and cytosine. In RNA uracil is found
instead of thymine. Chemical analysis also showed that polynucleotide chains
of RNA contain ribose sugar instead of deoxyribose (Fig .9.8).

Fig: 9.8: Chemical structure of RNA.

The self-complementary sequences in the RNA strand base pair with bases
present in the other strand (intrachain base-pairing) which results in folding of
the ribonucleotide chain into complex structural forms consisting of bulges and
helices (Fig.9.9). The three-dimensional structure of RNA is critical for its
stability and function, allowing the ribose sugar and the nitrogenous bases to
be modified in numerous different ways by cellular enzymes that attach
chemical groups (e.g., methyl groups) to the chain. Such modifications
facilitate the formation of chemical bonds between distant regions in the RNA
strand, leading to complex contortions in the RNA chain, which further
stabilizes the RNA structure. RNAs can also form complexes with molecules
known as ribonucleoproteins (RNPs).

(a) (b)
Fig 9.9: a) Comparison of DNA and RNA; b) Single strand of RNA. 19
Block 3 DNA-Blueprint of Life
Depending upon their presence in the plant and nature, RNA is of two types,
namely genetic RNA and non-genetic RNA.

Genetic RNA- In most of the plant viruses, RNA is the genetic material. Such
RNA contains information which is normally found in DNA. In other words,
RNA has replaced DNA in such cases. The RNA present in these viruses
could be single stranded or double stranded. In most of the bacteriophages,
RNA is the genetic material (see Table 9.1). Viroids and virusoids are other
two classes of small RNAs (~350 bases) found in plants. Viroids function
independently without encapsidation by a protein coat, while Virusoids (also
called satellite RNAs) are encapsidated by plant viruses packaged together
with a viral genome. Virusoids cannot replicate independently, and need an
association with the virus. The double stranded RNA follows the same rules of
base pairing as DNA.

Non-genetic RNA- In some organisms, genetic information is contained in

RNA which gets transmitted only through DNA. Although RNA is present in
good quantity, it does not serve as a genetic material. This type of RNA is
known as non-genetic RNA. Such a non-genetic RNA is synthesized on DNA
template.

The non-genetic RNA species found in cells for specific functions include: i)
Anti-sense RNA also called mic RNA (messenger RNA inhibiting
complementary RNA) is synthesized sometimes on the strand complementary
to the one used for mRNA synthesis. This is used for regulation of DNA
synthesis and gene expression both in vivo and in vitro; ii) HnRNA is
synthesized from split genes in eukaryotes.

Table 9.1: Different RNA viruses and the nature of RNA associated with
them.

Virus Types of RNA

Plant Viruses
Mosaic virus (MV) Single stranded
Wound Double stranded
Animal Viruses
Influenza virus Single stranded
Rous sarcoma Single stranded
Poliomyelitis Single stranded
Reovirus Double stranded
Bacteriophages
MS2, F2, r17 Single stranded

On the basis of functioning, RNA has been classified into three types:

• Messenger RNA (mRNA),

• Transfer RNA (tRNA),

20 • Ribosomal RNA (rRNA)

Unit 9 Genetic Material
Messenger RNA (mRNA) - This type of RNA carries genetic information
contained in DNA from the DNA in the nucleus to ribosomes (sites of protein
synthesis or translation). It constitutes a small fraction (5% - 10%) of total
RNA present in the cell. It is synthesized in the cell nucleus and then
transported out of the cell to facilitate protein synthesis and code sequencing
on proteins. This type of RNA is rather short-lived. The molecular weight of
this RNA ranges from 500 to 2000 kDa.

Ribosomal RNA (rRNA) - This is the most stable kind of RNA and is
associated with the ribosomes. They constitute about 60 to 80% of the total
RNA is the cell. rRNA is the part of the ribosomes. It catalyzes the synthesis of
proteins. It plays a major part in the binding to mRNA, recruitment of tRNA,
and catalysis of peptide bond formation between amino acids.

Transfer RNA (tRNA) - This is also known as soluble RNA (sRNA). It makes
another small fraction (10-15%) of RNA. It is a small RNA chain of about 80
nucleotides. These are the smallest molecules of RNA and work as adapter
molecules for carrying amino acid molecules to the site of protein synthesis.
The non-coding RNA molecule acts as the intermediate between nucleotide
and amino acid sequences.All these three types of RNA are found in all
organisms (Fig. 9.10).

Fig 9.10: A clover-leaf model of t RNA.

Apart from above described RNA some other RNA molecules are also found in
the cell.
Small Nuclear RNA (snRNA) mediates the processing of primary transcripts
(Protein sequences) in the nucleus to produce functional elements which then
are exported to the cytosol during DNA transcription.
Small nucleolar RNA (snoRNA) are small RNAs (about 60-300 nucleotides)
found in the cell nucleolus that play a role in the synthesis of ribosomes. 21
Block 3 DNA-Blueprint of Life
MicroRNAs (miRNAs) is a small non-coding RNA that is single-stranded,
containing only 22 nucleotides. These are found in plants, all animals, and
some viruses and play a role in RNA silencing and post-transcriptional gene
expression regulation.
You can comprehend the basic differences between DNA and RNA by going
through Fig. 9.11.

Fig.9.11: Comparison of DNA and RNA structure.

SAQ 2
a) State whether following statements are True or False:

i) The four bases present in RNA, are adenine, guanine, uracil and
cytosine.

ii) RNA contain ribose sugar instead of deoxyribose.

iii) RNAs are unable to form complexes with protein molecules known
as ribonucleoproteins (RNPs).

iv) Viroids and virusoids are other two classes of small RNAs (~350
bases) found in plants.

v) In most of the plant viruses, RNA is the genetic material.

b) Draw a fine structure of RNA.

c) Write the full form of the following:

i) snRNA

ii) snoRNA

iii) miRNA
22
Unit 9 Genetic Material

9.5 TRANSFORMATION EXPERIMENTS FOR

CONFIRMATION OF DNA AS GENETIC
MATERIAL
By the early twentieth century, scientists knew that chromosomes contain
genes, the bearers of hereditary information. They also knew that
chromosomes are made of DNA and proteins. But it took the first fifty years of
the twentieth century and many scientists to discover that DNA is the genetic
material. The evidence for this is given below. A series of experiments were
conducted in the 1920s which finally established that DNA is the genetic
material. We are going to describe these experiments below.

9.5.1 Griffith’s Experiment

You already know that Fredrich Miescher in 1898 isolated an acidic chemical
from nuclei of pus cells which he termed as nuclein. In 1928, British physician
Frederick Griffith while studying the pathogenic strain of bacterium called
pneumococcus (causes pneumonia) discovered that the bacterium
(Streptococcus pneumoniae) exists in two forms called S strain and the R
strain. The S strain produces smooth and shiny colonies when grown on solid
agar medium. This is because of mucous, polysaccharide coat secreted by the
cell. The R strain lacks the ability to produce mucous coat, therefore produces
colonies which are rough. When the S strain is injected into mice, the
bacterium triggers a fatal pneumonia. The disease is caused because S strain
had a polysaccharide coat which protects the bacterial cell from attack by the
mouse’s immune system. When R strain is injected into mice, the bacterium is
not able to cause pneumonia. He suggested that since R stain lacks
polysaccharide coat hence bacterial cell gets attacked by mouse’s immune
system. The injecting of live R- strain or dead S-strain alone did not caused
disease in mice.

Fig 9.12: Griffith’s bacterial transformation experiment. 23

Block 3 DNA-Blueprint of Life
The injection of heat killed S strain did not induce disease. The pneumonia
was caused when mice was injected with a mixture of live R strain and dead S
strain. Griffith autopsied animals that were injected with the mixture of live R
and dead S strain. He concluded that non pathogenic strain somehow
converted into pathogenic S strain by a substance present in the heat killed S
strain. He called the phenomenon as genetic transformation (Fig. 9.12). The
active substance responsible for inducing pathogenicity was termed as
‘transforming principle’. The chemical factor from heat killed S strain was
able to cause heritable change of non-pathogenic R stain into pathogenic S
strain.

9.5.2 Avery’s Experiments

Later on Oswald Avery, Colin McLeod and Maclyn Mc Carty in 1944 pursued
further investigations and provided evidence that the transforming substance
in heat killed S strain of Pneumococcus was nucleic acid of DNA and not
something residing in the cell wall. They separated and tested the molecules
found in the debris of the dead S cells for transforming ability one by one. The
tests showed that the polysaccharides themselves do not transform the rough
cells. Therefore, the polysaccharide coat, although undoubtedly concerned
with the pathogenic reaction, is only the phenotypic expression of virulence.
Avery et al. found that only one class of molecules, DNA, induced the
transformation of R cells. They deduced that DNA is the agent that determines
the polysaccharide character and hence the pathogenic character. This gave
the first confirmation and evidence that DNA carries the genetic information
and is the main hereditary material found in living organisms.

9.5.3 Hershey-Chase Bacteriophage Experiment

Bacteriophages are the viruses that infect bacteria. Bacteriophage T2 infects
bacterium Escherichia coli (E. coli). During infection, the virus attaches to the
bacterial cell surface and injects material into the cell. The bacterial cell begins
to produce thousands of new copes of the virus. The material injected into the
bacterial cell carries the genetic information that guides the production of the
virus. In 1952, Alfred Hershey and Martha Chase designed an experiment to
know that T2 virus is made up of two kinds of molecules DNA and proteins
(Fig 9.13). The proteins of the T2 virus contain elemental sulfur (present in
amino acids methionine and cysteine) and not phosphorus, while the DNA
contains phosphorus (present as sugar phosphate backbone) but not sulfur.
They prepared two batches of T2 phage particles with different radioactive
labelling. In one batch the phage proteins were labelled with the radioactive
isotope 35S and in the other batch the phage DNA was labelled with isotope
32
P. By radioactive labelling Hershey and Chase were able to trace the fate of
protein and DNA during infection process.

They started the experiment by mixing the radioactive phage with intact
bacterial cells. They allowed phage particles to attach to the bacterial cell
surface and let them inject their genetic material into the cells. Then they
removed protein coats from the surface of the bacterial cells by agitating the
suspension in a blender and recovered the bacterial cells by centrifugation.
They measured radioactivity in the supernatant liquid and in the pellet of
bacteria at the bottom of the tube. The studies showed that most of the (65%)
24 of the 32P remained in the bacterial cells while the bulk of the 35S was released
Unit 9 Genetic Material
in the surrounding medium. Since 32P labelled the viral DNA and 35S labelled
the viral protein, they concluded that DNA had been injected into the bacterial
cells and functions as the genetic material of phage T2.

It is important to know that when the 32P-labelled phages were used in the
experiment, the majority of the radioactivity was found inside the bacterial
cells, which indicates that the phage DNA has entered the cells. This 32P can
also be recovered from phage progeny. But when the 35S-labelled phages
were used in the experiment, most of the radioactive material was found in the
phage ghosts, thus it indicates that the phage protein had never entered the
bacterial cell. Thus it was concluded that DNA is the hereditary material and
the phage proteins are simply structural packaging which is discarded after
delivering the viral DNA to the bacterial cell.

When infected radioactively labelled bacteria were suspended in fresh liquid

and incubated for longer time, the 32P was transferred to some of the offspring
phage particles. This gave the view that genes are made up of DNA.

Fig 9.13: Experiment of Hershey and chase to demonstrate DNA as the

hereditary material. a) The protein coat is labelled with radioactive
35
sulfur ( S), not found in DNA. b) The DNA is labelled with radioactive
32 32
phosphorus ( P), not found in protein. Only the P is injected into the
E. coli, indicating that DNA is the agent necessary for the production of
new phages.

Apart from experimental evidence, biochemical evidence due to certain facts

such as those given below confirmed that DNA is the genetic material.

i) Amount of DNA in every cell of an organism is the same.

ii) Amount of DNA in gametes is half that of DNA present in somatic cells.

iii) Amount of DNA is the same in all members of same species. 25

Block 3 DNA-Blueprint of Life
iv) Amount of DNA in a cell is constant and does not change by internal or
external environment.

v) DNA or amount of genetic information is proportional to evolutionary

complexity.

SAQ 3
a) State whether following statements are ‘True’ or ‘False’ for Griffith
experiment.

i) In his experiment he discovered that the bacterium (Streptococcus

pneumoniae) exists in two forms called S strain and the R strain

ii) In his experiment he found that R strain lacks the ability to produce
mucous coat but can produce pneumonia.

iii) In his experiment he found that S strain has the ability to produce
mucous coat but was unable to produce pneumonia.

iv) In his experiment he found that injecting of live R- strain or dead S-

strain alone did not caused disease in mice.

b) Only through diagram show Hershey-Chase Bacteriophage experiment.

9.6 SUMMARY
• DNA acts as the genetic material in majority of living organisms except in
some viruses where RNA is present as the main hereditary material. In
some organisms such as bacteriophages, both DNA as well RNA carry
genetic information for the cells.

• Nucleic acid molecule is a long chain polymer (Polynucleotide)

composed of monomeric units called nucleotides. These monomer units
of nucleotide are jointed by the formation of phosphodiester bonds. Each
nucleotide consists of three components - nitrogen-containing bases, a
five-carbon sugar, and a phosphate group. A phosphate group and
nitrogen containing aromatic base is attached to five carbon sugar. The
nitrogenous base is either a purine or a pyrimidine.

• DNA is a double helical structure with two helices of the nucleotide

chains coiled around each other spirally to form double helix. The
structure of the double helix is like a ladder, with base pairs running
through the middle like rings and sugar and phosphate molecules along
the outside.

• Levene was the first to discover the order of the three major components
of a single nucleotide (phosphate-sugar-base), carbohydrate component
of DNA and RNA. He proposed that nucleic acids were composed of a
series of nucleotides. Watson and Crick determined the structure of
deoxyribonucleic acid (DNA). Rosalind Franklin and Maurice Wilkins
26 used X-ray diffraction technique to deduce the structure of DNA.
Unit 9 Genetic Material
• RNA is a single stranded molecule. It consists of ribose nucleotides
attached by phosphodiester bonds. The four bases present in RNA are
adenine, guanine, uracil and cytosine. Three types -messenger RNA
(mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA) are present
in all organisms.

• The confirmation of DNA as genetic material came from the experiments

conducted by Frederick Griffith using the pathogenic strain of bacterium
called Streptococcus pneumoniae and Hershey and Chase using
bacteriophages particularly bacteriophage T2 that infects bacterium
Escherichia coli (E.coli).

• Biochemical evidences also suggested that DNA is the genetic material.

9.7 TERMINAL QUESTIONS

1. Describe briefly the history of discovery of DNA.

2. What is the Chargaff rule? Discuss its importance.

3. Differentiate between genetic and non genetic RNA.

4. Describe the first experiment which confirmed that DNA carries genetic
information.

5. How did Avery, McLeod and McCarty’s experiment on bacterial

transformation provide evidence for DNA being the genetic material?

6. Write short notes on the following:

i) Types of RNA based on functioning

ii) Structure of Nucleic acids

iii) Genetic RNA.

9.8 ANSWERS
Self-Assessment Questions
1. a) i) 4

ii) Chromosomes

iii) inosinic acid

iv) nuclein

b) A=Adenine G= Guanine, C= Cytosine and T= Thiamine

2. a) i) True; ii) True; iii) False; iv) False; v) True

b) Refer to Fig.9.10

c) i) snRNA =Small Nuclear RNA

ii) snoRNA= Small nucleolar RNA

iii) miRNA =MicroRNAs 27

Block 3 DNA-Blueprint of Life
3. a) i) True; ii) False; iii) False; iv) True

b) Refer to Fig 9.13

Terminal Questions
1. Refer to Subsection 9.3.1

2. Refer to Subsection 9.3.2

3. Refer to Section 9.4

4. Refer to Subsection 9.5.1

5. Refer to Subsection 9.5.2

6. i) Refer to Section 9.4

ii) Refer to Section 9.3.2 and 9.4

iii) Refer to Section 9.4

Acknowledgement
Fig.9.4 : Source:
https://d2cbg94ubxgsnp.cloudfront.net/Pictures/480xAny/6/3/1/8
4631_feature-dna_fig2_300.jpg

Fig. 9.5 : Source:

https://lh3.googleusercontent.com/proxy/VPBqjLBFhJg2Tc59r-
ebeZqLaNc-

Fig. 9.10 : Source:

https://www.google.co.in/imgres?imgurl=https%3A%2F%2Fcdn.l
ecturio.com%2Fassets%2FTransfer-RNAs-tRNA-
e1614263850807-1200x709.png&imgrefurl=

Fig.9.11 : https://www.google.co.in/imgres?imgurl=https%3A%2F%2Fcdn.
technologynetworks.com%

Fig. 9.12 : https://www.google.co.in/imgres?imgurl=https%3A%2F%2Fdigit

alcommons.rockefeller.edu%2Ftransforming-principle-dna%

Fig 9.13 : From: W. H. Freeman; 2000.DNA: The genetic material, An

Introduction to Genetic Analysis. 7th edition. New York:

28