History of DNA

The human hereditary material known as

deoxyribonucleic acid, or DNA, is a long molecule
containing the information organisms need to
both develop and reproduce. DNA is found in
every cell in the body, and is passed down from
parent to child.

Although the discovery of DNA occurred in 1869 by

Swiss-born biochemist Friedrich Miescher, it took
more than 80 years for its importance to be fully
realized. And even today, more than 150 years
after it was first discovered, exciting research and
technology continue to offer more insight and a
better.

● 1866 — Before the many significant discoveries

and findings, Gregor Mendel, who is known as
the “Father of Genetics,” was actually the first
to suggest that characteristics are passed
down from generation to generation. Mendel
coined the terms we all know today as
recessive and dominant.
● 1869 — Friedrich Miescher identified the “nuclein”
by isolating a molecule from a cell nucleus that
would later become known as DNA.
● 1881 — Nobel Prize winner and German biochemist
Albrecht Kossel, who is credited with naming DNA,
identified nuclein as a nucleic acid.
● Early 1900s — Theodor Boveri and Walter Sutton
were independently working on what’s now known
as the Boveri-Sutton chromosome theory, or the
chromosomal theory of inheritance.
● 1944 — Oswald Avery first outlined DNA as the
transforming principle, which essentially means
that it’s DNA, not proteins, that transform cell
properties .
● 1944-1950 — Erwin Chargaff discovered that DNA is
responsible for heredity and that it varies between
species. His discoveries, known as Chargaff’s
Rules, proved that guanine and cytosine units, as
well as adenine and thymine units, were the same
in double-stranded DNA, and he also discovered
that DNA varies among species.
● 1953 — Watson and Crick published on DNA’s
double helix structure that twists to form the
ladder-like structure we think of when we picture
DNA.
 DNA

Adenine Thymine

Guanine Cytosine
 Components of DNA
They are major components of all cells. DNA is
found predominantly in the nucleus while RNA is
predominant in cytoplasm. DNA is the genetic
material of most organisms including many
viruses. Some viruses,’ however, have RNA as their
genetic material.

(i) Phosphoric Acid:

The phosphoric acid (H3PO4) in the nucleic acid is
called phosphate . Phosphoric acid has three
reactive hydroxyl (-OH) groups of which two are
involved in forming sugar phosphate backbone of
DNA. The phosphate makes a nucleotide negatively
charged.

(ii) Sugar Molecule:

DNA contains a five carbon sugar, hence it is a
pentose sugar. Since one oxygen atom at the 2′
carbon is missing, hence it gets its name
2′-deoxyribose. Four of the five carbon atoms plus a
single oxygen atom forms a five-membered ring.
The fifth carbon atom is outside the ring and forms
a part of a -CH2 group.
 Organic Bases:

(a) Pyrimidine:
Pyrimidine bases are made up of a six-membered
pyrimidine ring which is similar to the benzene
ring except that it contains nitrogen in place of
carbon at the positions 1 and 3. Pyrimidine bases
are of 2 types — thymine and cytosine, commonly
abbreviated as T and C respectively.

(b) Purine:
It is a derivative of pyrimidine. It consists of a
pyrimidine ring and a five- membered imidazole
ring (having nitrogen at 7 and 9 positions) which
are fused together at 5 and 4 positions. There are
two purine compounds namely – adenine (A) and
guanine (G).
 Molar Ratio of Nitrogenous Bases
in DNA (Chargaff Rules, 1955):
(i) The purine and pyrimidine components occur in
equal amounts in a DNA molecule.
(ii) The amounts of adenine (A) is equivalent to the
amount of thymine (T) and the amount of cytosine
is equivalent to that of guanine (G).
(iii) In DNA, A + G / T + C value is always one or
nearly one.
(iv) The base ratio A +T/G + C may vary in the DNA of
different groups of organisms but is constant for
particular species. Therefore, this ratio has been
used to identify the DNA from a particular species.
 Nucleoside:

A base combined with a sugar molecule is

called a nucleoside. When deoxyribose sugar
binds with base, it makes deoxyribonucleoside.
Obviously, in DNA four different nucleotides are
found.
These are:
(i) Deoxycytidine
(ii) Deoxythymidine
(iii) Deoxyadenosine
(iv) Deoxyguanosine
Nucleotide:
A nucleotide is derived from a nucleoside by
addition of a molecule of phosphoric acid. The
phosphate molecule is linked with sugar
molecule at carbon number 5 or at carbon
number 3.
The nucleotides in DNA are of 4 types:
(i) Deoxycytidylic acid
(ii) Deoxythymidylic acid
(iii) Deoxyadenylic acid
(iv) Deoxyguanylic acid

Structure of four different 5’p 3’OH nucleotides found in DNA.

 James Watson and Francis Crick Model

The salient features of the Double-helix structure of

DNA are as follows:

(i) It is made of two polynucleotide chains, where the

backbone is constituted by sugar-phosphate, and the
bases project inside.

(ii) The two chains have antiparallel polarity. It

means, if one chain has the polarity 5'→3' , the other
has 3'→5' .
(iii) The two chains are coiled in a right-handed
fashion. The pitch of the helix is 3.4 nm (a
nanometre is one billionth of a metre, that is 10-9
m) and there are roughly 10 bp in each turn.
Consequently, the distance between a bp in a helix
is approximately 0.34 nm.
(iv) The bases in two strands are paired through
hydrogen bond (H-bonds) forming base pairs (bp).
Adenine forms two hydrogen bonds with Thymine
from opposite strand and vice-versa. Similarly,
Guanine is bonded with Cytosine with three
H-bonds.

(v) The plane of one base pair stacks over the

other in double helix. This, in addition to H-bonds,
confers stability of the helical structure.
 Forms of DNA:
The DNA molecules exhibit a considerable amount
of conformational flexibility. It can exist in A, B, C, D
and Z forms.

A form (A-DNA) has 11 base pairs. The base pairs are

considerably tilted from the axis of the helix. The
axial rise is 2.56Å. The helix observed under
conditions of dehydration and high concentrations
of salt is wider and shorter than B- helix, the
distinction between the major and minor grooves
are reduced.

B form (B-DNA) is the structure proposed by Watson

& Crick and is the native conformation of DNA in
solution.

C form (C-DNA) results by reduction of hydration of

the B form below 66% with excess of salt still
present. The size of helix of C form DNA is greater
than Å type of DNA but is smaller than B-DNA. It is
about 31 Å. There are 9.33 base pairs per turn. The
axial rise of base pairs is 3.32 Å with a tilting of
about 7.8°.
D form (D-DNA) and E form (E-DNA) are found rarely
as extreme variants. In case of D- form there are 8
base pairs per turn of helix. An axial rise of base
pairs is 3.03 Å with tilting of about 16.7°.In case of
E-form, there are 7.5 base pairs per turn of helix.

Z form (Z-DNA) is an unique left-handed double

helical form with a zig-zag sugar-phosphate
backbone in antiparallel organization. This DNA has
been called Z-DNA.

Characteristics of the different form of DNA

 Nucleosome

The nucleosome model proposed by R.D. Kornberg in

1974 is the most significant one that explains the
structure of nucleosomes. This model was confirmed
and named by P. Oudet in 1975. According to this
nucleosome model, the histone particle is wrapped
with DNA around it.

Nucleosome Model of Chromosome:

A chromosome molecule contains many nucleosomes
that are repeated. Each nucleosome is made up of
1.65 loops of DNA wrapped around 8 histone
proteins. A nucleosomes wrapped DNA molecule
contains approximately 146 base pairs.
The nucleosome contains two copies of each of the
four nucleosome histones – H2A, H2B, H4 and H3.
Histone octamers are the collective name for the
eight histones.
Histones proteins are positively charged molecules
that can form stronger bonds with negatively charged
DNA molecules.

The DNA that wraps around the histone structure is

known as the core or wrapping DNA.

Each nucleosome is a disc-shaped particle with a

height of about 5.7 nm and a diameter of about 11 nm.
These repeated bead-like structures are linked by a
linker DNA molecule, which typically consists of 54
base pairs and an H1 histone protein. As a result, the
H1 histone binds to the location where DNA exits and
enters the nucleosome. The H1 histone does not
belong to the histone octamer group.
The nucleosomes repeat at approximately 200 base
pairs or nucleotide intervals on average.
 Early Experiments on DNA
Transformation Experiment:

Transformation experiment was initially conducted by

F. Griffith in 1928. The injected a mixture of two strains
of Pneumococcus (Diplococcus pneumoniae) into
mice. One of these two strains, S III was virulent and
other strain R II was non-virulent (causing no
infection).

Heat-killed virulent S III strain when injected, showed

that infectivity after heat killing is lost. The mice
injected with a mixture of R II (living) and S III (heat
killed) died and virulent Pneumococcus could be
isolated from these mice. This phenomenon was
described as transformation.

O. T. Avery, C. M. Macleod and M. McCarthy repeated

Griffith’s experiment in an in vitro system in order to
identify the transforming principle responsible for
converting non-virulent into virulent type and
reported their results in 1944. Virulence in
Pneumococcus depends on a polysaccharide capsule
which is present in virulent strain S III and is absent in
non-virulent strain R II.
The cells of non-capsulated type – Rll were treated
with an extract of DNA from capsulated strain S III. A
few cells of S III type could be isolated from the
mixture.

This phenomenon of transferring characters of one

strain to another by using a DNA extract of the
former is called transformation. When the extract
was treated with DNAse (an enzyme which destroys
DNA) this transforming ability was lost. Proteases
(enzymes which destroy proteins) did not affect the
transforming ability. These experiments thus
indicated that DNA and not the protein, is the
genetic material.
Hershey and Chase Experiment

In 1952 A. Hershey and Chase conducted

experiments on Bacteriophage and E. Coli

They took two groups of Bacteriophage-

● Radio labeled 32P DNA bacteriophage

● Radio labeled 32S capsid bacteriophage

The experiment included 3 steps:

1. Infection- bacteriophage are allowed to attack E.

Coli separately.
2. Blending- The capsid or protein coat was
removed from bacteria by a blender.
3. Centrifugation- Virus particle were separated
from bacteria by spinning them in centrifuge
machine.

After centrifugation, it was found that the bacteria

which was attacked by bacteriophage having a
Radioactive DNA, contains Radioactive substance
within it.
It means that DNA has transferred from
bacteriophage to the bacteria and not the proteins.
Thus, DNA is regarded as the genetic material.
 Semi Conservative method of
replication
Semi conservative method usually occurs during
S-phase of cell cycle when chromosomes are in
highly extended form. As proposed by Watson and
Crick, DNA replication is semiconservative (a type of
replication in which one strand of the daughter
duplex is derived from the parent while the other
strand is formed anew).

Semi-conservative replication of DNA was proved by

the work of Matthew Meselson and Franklin Stahl
(1958). They grew Escherichia coli for many
generations in a medium having heavy isotope of
nitrogen, in the form of 15NH4Cl, till the bacterial DNA
became completely labelled with heavy isotope.

The labelled bacteria were then shifted to fresh

medium having normal or 14N nitrogen. Samples
were taken for each generation (one generation
takes 20 minutes as E. coli divides in 20 minutes)
and the DNA was tested for the heavy isotope of
nitrogen through density gradient centrifugation
using caesium chloride. Caesium chloride is highly
water soluble heavy salt.
When spun in centrifuge at high speed (say 50,000
revolutions per minute) the salt forms a density
gradient with heaviest most concentrated region at the
bottom and successively less concentrated lighter one
towards the surface. When DNA is mixed with caesium
chloride it will settle down at a particular height in
centrifugation, heavier towards the base and lighter one
higher up.

Fluoro-chrome called ethidium bromide is used to

enhance contrast as the fluorochrome is specific for
DNA. Meselson and Stahl found that DNA of the first
generation was hybrid or intermediate (15N and 14N). It
settled in caesium chloride solution at a level higher
than the fully labelled DNA of parent bacteria (15N15N).
The second generation of bacteria after 40 minutes
contained two types of DNA, 50% light (14N14N) and 50%
intermediate (15N14N).

The third generation of bacteria after 60 minutes

contained two types of DNA, 25% intermediate (15N14N)
and 75% light (14N14N) in 1 : 3 ratio. The fourth
generation after 80 minutes contained 12.5% 15N14N
and 87.5% 14N14N DNA in 1 : 7 ratio.
This observation is possible only if the two strands of
DNA duplex separate at the time of replication and
act as a template for the synthesis of new
complementary strands of DNA having normal or 14N.
This will produce two DNA duplexes with one old
strand (15N) and one new strand (14N).

During the formation of second generation, 15N and

14N strands of DNA duplex separate to function as
templates so that 50% of new DNA duplexes possess
only normal or l4N strands while another 50% have
both 15N and 14N strands (Figs. 6.11& 6.12). In this way
at each replication, one strand of parent DNA is
conserved in the daughter while the second is freshly
synthesised
 Conclusion:

DNA is very important for life. It can replicate well,

which means that the next generation will retain the
characteristics of the parents. It is capable of change,
which means that it provides for variation and was
crucial for evolution to occur. It also codes for
proteins that help express genes and traits of the
organism.
 ACKNOWLEDGEMENT

I extend my heartfelt gratitude to my biology teacher

Mr.Udayanath Sahoo for his help and cooperation
without his support and valuable suggestion it would
have been impossible on my part to complete the
project.

I extend my special thanks to our respective Principal

Mr. S.K.Bhoi for his cooperation and encouragement.

I am thankful to CBSE Authorization for their idea

about the project for it which helped me to come
across many facts about which I don't know.

Name:
Class: XII (SCIENCE)
Roll no:
 Certificate

This is to certify that ,

a student of Class XII (Science) of D.A.V Public
School M.C.L Kalinga Area has successfully
completed the ‘Study of DNA structure and
Experiments’ under the of Mr. Udayanath Sahoo
(HOD, Biology) as per requirement of
CBSE-AISSCE-12.

Mr. Udayanath Sahoo Mr. S.K Bhoi

PGT (Biology) (Principal)
DAV Public School DAV Public School
MCL, Kalinga Area MCL, Kalinga Area
Signature of Internal Signature of external
 DNA
Study of DNA structure
and Related
Experiments