University of Zakho

General Science Department

Report title
DNA is Genetic Material

Supervisor:
Prepared by:
Group:A

2019-2020

Abstract
DNA (DNA = deoxyribonucleic acid)

DNA is the genetic material of all living cells and of many viruses.

DNA is: an alpha double helix of two polynucleotide strands.

The genetic code is the sequence of bases on one of the strands.

The chromosomes contain 90% of the cell’s DNA.

10% is present in mitochondria and chloroplasts.

Adenine (A) and Guanine (G) are purine bases

Thymine (T) and Cytosine (C) are pyrimidine bases

Hydrogen bonds link the complementary base pairs:

 Two between A and T (A = T)

 Three between G and C (G ≡ C)
A single unit in the chain is a nucleotide.
 This consists of a phosphate group,
 a pentose sugar (D = DNA; R = RNA) and
 an organic base (ATGC = DNA; AUGC = RNA)

DNA is genetic material:

1928: Griffith - Transforming Principle

1944: Avery, MacLeod & McCarty - DNA is Transforming Principle

1950: Chargaff’s Rules - A=T, G=C

1952: Hershey & Chase - Blender Experiment

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1953: Watson & Crick - A Structure for DNA

Table of Contents:

Introduction 5

Griffith and the Transforming Principle 6- 7

The Experiments of Avery, MacLeod & 8- 9

McCarty

Hershey–Chase experiment 10-13

DNA and Chromosome 14- -17

18- 23
DNA Structure

X-ray Diffraction Patterns Produced by DNA fibers 24

25
Erwin Chargaff’s Experiment
26- 27
References

List of Figures

Figure 1. The experiment of Griffith that demonstrated the concept of

the transforming principle. 6

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Figure9.3. The Experiments of Avery, MacLeod & McCarty 9

Figure4.4.Bacteriophages and E.Coli 13

Figure2:DNA and Chromosome 17

Fig. 3. The single strand structure of DNA 21

Fig.6. Watson and Crick’s proposed structure 22

Fig.38.4.Watson and Crick’s double helical molecule 23

Fegure.1.x ray Diffracted by DNA 24

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 Introduction
DNA is the cornerstone of genetics and is the perfect place to start for an
introduction to genetics. DNA stands for deoxyribonucleic acid and it is the
molecule that holds the genetic information for a cell and an organism.
DNA - introduction to geneticsA DNA molecule contains a code that can be used
by a cell to express certain genes. Specific sections of a DNA molecule provides
the information to build specific proteins which can then be used by a cell to
express the desired gene.
A DNA molecule is a nucleic acid, one of the four molecules of life. It comes in the
form of a long, linear molecule referred to as a strand. Each strand of DNA is
bonded to a second strand of DNA to form a DNA double helix. In eukaryotic cells,
DNA is found in the nucleus as a tightly coiled double helix.
DNA molecules are replicated during cell division. When a cell divides, the two
new cells contain all the same DNA that the original cell had.
In sexual reproduction with two parents, half of the DNA of the offspring is
provided by each of the parents. The genetic material of a child is made from 50%
of their mother’s DNA and 50% their father’s DNA.

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 Griffith and the Transforming Principle

The Concept

The experiments of Griffith and Avery, MacLeod and McCarty are closely
related.

Griffith developed the concept of the transforming principle. The prinicple was

able to transform a non-pathogenic bacteria into a pathogenic strain. Changing

phenotype is one of the characteristics of the hereditary material. Griffith called

the factor that changed the phenotype the tranforming principle. Avery, and
MacLeod performed a series of experiments that demonstrated the hereditary

McCarty, materials was DNA.(1)

Figure 1. The experiment of Griffith that demonstrated the concept of the

transforming principle.

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Griffith was able to show that if you heat kill a Type IIIS strain and injected it into
the mouse, the mouse lived. But if you mixed the heat-killed type IIIS material
with live type IIR bacteria, the mouse would die. Furthermore, the autopsy
showed that the mouse became infected with the Type IIIS strain. These meant
that some material from the Type IIIS strain was taken up by the Type IIR strain to
convert it into the Type IIIS strain. Griffith termed the material the transforming
principle.

One feature of the genetic material is its ability to control phenotype. In Griffith's
experiment, the bacterial strains have several phenotypes. The R types are not
only non-lethal, and they have a rough (R) appearance on a blood agar plate. The
S type are distinct from the R type: they are lethal and have a smooth morphology
on the plates. The S types have a polysaccharide capsule that is lacking in the R
types. Each capsule type is distinguished using antibodies; the type II capsule is
antigenically distinct from the type III. The trans- formation from type II to type III
and the conversion of type R to S are each distinct phenotypic changes.(1)

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 The Experiments of Avery, MacLeod & McCarty
 realized that Griffith’s observations could be used to identify the genetic
material or “transforming principle”
 In essence, the formation of the capsule is guided by the bacteria’s genetic
material.
– Transformed bacteria acquired information to make the
Capsule.
– Variation exists in ability to make capsule
– The information required to create a capsule is replicated and transmitted
from mother to daughter cells
 At the time of their experimentation in the 1940s, it was known
that DNA, RNA, proteins and carbohydrates are the major
constituents of living cells.
 Prepared cell extracts from smooth cells (IIIS) and added to
rough cells (IIR) for transformation in culture medium
 Only the DNA enriched extract was able to convert rough cells
into smooth cells.(2)

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9
 Hershey–Chase experiment

The Hershey–Chase experiments were a series of experiments conducted in

1952[3] by Alfred Hershey and Martha Chase that helped to confirm that DNA is
genetic material. While DNA had been known to biologists since 1869,[3] many
scientists still assumed at the time that proteins carried the information for
inheritance because DNA appeared to be an inert molecule, and, since it is
located in the nucleus, its role was considered to be phosphorus storage. In their
experiments, Hershey and Chase showed that when bacteriophages, which are
composed of DNA and protein, infect bacteria, their DNA enters the host bacterial
cell, but most of their protein does not. Hershey and Chase and subsequent
discoveries all served to prove that DNA is the hereditary material.

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 Methods and results

Hershey and Chase needed to be able to examine different parts of the phages
they were studying separately, so they needed to distinguish the phage
subsections. Viruses were known to be composed of a protein shell and DNA, so
they chose to uniquely label each with a different elemental isotope. This allowed
each to be observed and analyzed separately. Since phosphorus is contained in
DNA but not amino acids, radioactive phosphorus-32 was used to label the DNA
contained in the T2 phage. Radioactive sulfur-35 was used to label the protein
sections of the T2 phage, because sulfur is contained in protein but not DNA.

Hershey and Chase inserted the radioactive elements in the bacteriophages by

adding the isotopes to separate media within which bacteria were allowed to
grow for 4 hours before bacteriophage introduction. When the bacteriophages
infected the bacteria, the progeny contained the radioactive isotopes in their
structures. This procedure was performed once for the sulfur-labeled phages and
once for phosphorus-labeled phages. The labelled progeny were then allowed to
infect unlabeled bacteria. The phage coats remained on the outside of the
bacteria, while genetic material entered. Disruption of phage from the bacteria by
agitation in a blender followed by centrifugation allowed for the separation of the
phage coats from the bacteria. These bacteria were lysed to release phage
progeny. The progeny of the phages that were labeled with radioactive
phosphorus remained labeled, whereas the progeny of the phages labeled with
radioactive sulfur were unlabeled. Thus, the Hershey–Chase experiment helped
confirm that DNA, not protein, is the genetic material.

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Hershey and Chase showed that the introduction of deoxyribonuclease (referred
to as DNase), an enzyme that breaks down DNA, into a solution containing the
labeled bacteriophages did not introduce any 32P into the solution. This
demonstrated that the phage is resistant to the enzyme while intact. Additionally,
they were able to plasmolyze the bacteriophages so that they went into osmotic
shock, which effectively created a solution containing most of the 32P and a
heavier solution containing structures called “ghosts” that contained the 35S and
the protein coat of the virus. It was found that these “ghosts” could adsorb to
bacteria that were susceptible to T2, although they contained no DNA and were
simply the remains of the original bacterial capsule. They concluded that the
protein protected the DNA from DNAse, but that once the two were separated
and the phage was inactivated, the DNAse could hydrolyze the phage DNA.(3)

Experiment and conclusions

Hershey and Chase were also able to prove that the DNA from the phage is
inserted into the bacteria shortly after the virus attaches to its host. Using a high
speed blender they were able to force the bacteriophages from the bacterial cells
after adsorption. The lack of 32P labeled DNA remaining in the solution after the
bacteriophages had been allowed to adsorb to the bacteria showed that the
phage DNA was transferred into the bacterial cell. The presence of almost all the
radioactive 35S in the solution showed that the protein coat that protects the
DNA before adsorption stayed outside the cell.(3)

Hershey and Chase concluded that DNA, not protein, was the genetic material.
They determined that a protective protein coat was formed around the

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bacteriophage, but that the internal DNA is what conferred its ability to produce
progeny inside a bacterium. They showed that, in growth, protein has no
function, while DNA has some function. They determined this from the amount of
radioactive material remaining outside of the cell. Only 20% of the 32P remained
outside the cell, demonstrating that it was incorporated with DNA in the cell's
genetic material. All of the 35S in the protein coats remained outside the cell,
showing it was not incorporated into the cell, and that protein is not the genetic
material.

Hershey and Chase's experiment concluded that little sulfur-containing material

entered the bacterial cell. However no specific conclusions can be made regarding
whether material that is sulfur-free enters the bacterial cell after phage
adsorption. Further research was necessary to conclude that it was solely
bacteriophages' DNA that entered the cell and not a combination of protein and
DNA where the protein did not contain any sulfur.(3)

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Figure4.4.Bacteriophages and E.Coli

DNA and Chromosome

DNA is the chemical form of storing the genetic information used in development,
function, and reproduction. It is a

macromolecule, composed of nucleotides as the monomer units. A nucleotide is

built with a nitrogen base and a

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phosphate group which is attached to a phosphate sugar. Four different
nitrogenous base types can be identified in

DNA: cytosine (C), guanine (G), adenine (A) and thymine (T). The phosphate group
of one nucleotide is attached to a

sugar of another nucleotide via phosphodiester bonds to form the sugar-

phosphate backbone. The polynucleotide

stands are joined together by hydrogen bonds forming between two complement
nitrogenous bases: A with T and C

with G. Hence, doublestranded DNA is formed, each strand is complementary to

the other strand. Also, the two

strands run in opposite directions, making the strands antiparallel. Double-

stranded DNA is coiled around each other

to form the DNA doublehelix.

DNA is organized into chromosomes for the easy package within the cell. The
sequence of four bases along the DNA

strand encodes the genetic information as genes. The size of the human genome
is 3.2 billion base pairs. Humans have(4)

about 21,000 of genes. More than 98% of human DNA is composed of noncoding
sequences whereas the other

sequences are encoded for proteins. A few differences between the gene
sequences make an individual identical.

During cell division, the exact replica of original DNA is synthesized by replication.

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A chromosome is the most organized structure of DNA. In eukaryotes, DNA
doublehelix is condensed with histone proteins to form nucleosomes.
Nucleosome structure is further coiled into a fiberlike structure called chromatin

fibers with a diameter of 250 nm. Chromatin is the normally existing form of DNA
within the nucleus. They exhibit a threadlike structure and are less condensed

compared to a chromosome. Chromatin is then further coiled to form

chromosomes. The diameter of a chromosome is 30 nm. Chromosomes can be
seen during the nuclear division even.

Eukaryotes consist of large, linear chromosomes whereas the prokaryotes consist

of a single, circular chromosome

condensed with histonelike proteins. Organization into chromosomes provides

the structural integrity to DNA

doublehelix. A chromosome consists of thousands of genes. The accessibility to

the sequence of the DNA at chromosomal level regulates the gene expression.
Humans have 46 individual chromosomes. There are 22 homologous pairs of
autosomes and 2 sex chromosomes. A chromosome also contains an origin of
replication,(4)

centromere, and telomeres. Metaphase chromosomes of a cell are used to

generate karyotypes for the analysis of chromosomal abnormalities

Main Difference – DNA vs Chromosome

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All known living organisms and many viruses utilize DNA as their genetic material
in order to store their genetic information. DNA is a bivalent structure that exists
as double helix. DNA doublehelix is condensed with histone proteins to

form a chromosome. The key difference between DNA and chromosome is that
DNA is the unorganized structure of the carrier of genetic information in most
organisms and chromosome is the most organized structure of DNA with

histones within a cell. Further, DNA stores the genetic instructions

whereas chromosomes allow the gene regulation of a DNA strand.(4)

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DNA Structure

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 Watson and Crick

Watson and Crick looked at previous structures proposed by Pauling and Corey

Consisted of…

Three intertwined chains

Phosphate near fibre axis
Bases on the outside

reasons it was unsatisfactory:

1) The material which gives the x ray diagrams is the salt not the free acid

2) Some of the Van der Waals distances appear to be too small.(5)

Watson and Crick looked at previous structure proposed by Fraser

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Consisted of:

 Another three chain

 Phosphates on the outside
 Bases on the inside

Reason it was unsatisfactory: It is ill defined and therefore could not be used

Watson and Crick’s Structure

 2 helical chains each coiled around the same axis

 Both chains are right handed
 The two chains are related by dyad perpendicular to fibre axis
 Each chain consists of phosphate diester groups
Joining β-d-deoxyribofuranose residues with 3-5 linkages
 Residues on each chain every 3.4 A
 36° between each adjacent residues in the same chain
 Structure repeats itself after 10 residues
 Open structure with high water content
 Bases tilt at low water content would compact it.(5)

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Watson and Crick’s Structure

Watson and Crick structure loosely resembles Furburg Model #1

 Bases were on the inside

 Phosphates were on the outside
 Sugar perpendicular to attached base.(5)

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Fig. 3. The single strand structure of DNA

Watson and Crick Novel Feature

The two chains are held together by Purine and Pyrimidine Bases

Planes of the bases is perpendicular to the fibre axis

Single base from one hydrogen bonded to the other

One must be purine and the other pyrimidine.(5)

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 The major importance of Watson and Crick’s proposed structure

It immediately suggests method for copying mechanism for genetic material

Fig.6. Watson and Crick’s proposed structure

Watson and Crick’s Conclusion:

X rays done of DNA so far are insufficient to

describe their structure proposed

Therefore…

Their structure must be regarded as unproved

until it has been checked against more exact

experimental results.(5)

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 Watson and Crick’s Acknowledgments

Within their paper, Watson and Crick stated that they were indebted to certain

individuals for assisting in their research:

Dr. Jerry Donohue for his advice and criticism on the model

- Specifically with respects to interatomic distances

Wilkins and Franklin for their published works(5)

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 X-ray Diffraction Patterns Produced by DNA fibers
Rosalind Franklin

 She used X-ray diffraction to study wet fibers of DNA

 The diffraction pattern she obtained suggested several structural features
of DNA.

– Helical

– More than one strand

– 10 base pairs per complete turn.(6)

Fegure.1.x ray Diffracted by DNA

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 Erwin Chargaff’s Experiment
 Chargaff pioneered many of the biochemical techniques for the isolation,
purification and measurement of nucleic acids from living cells
 It was already known then that DNA contained the four bases: A, G, C and T
 Chargaff analyzed the the base composition of DNA in different species to
see if there was a pattern.(7)

Chargaff’s rule
 Percent of adenine = percent of thymine (A=T)
 Percent of cytosine = percent of guanine (C=G)
 A+G = T+C (or purines = pyrimidines)(7)

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 References

(1) Doermann, A. H., 1995, Carnegie Institution of Washington Yearbook, No.

47, 49
(2) Satchwell SC, Drew HR & Travers AA (1986) Historical article:. J Mol Biol
191, 61–64.
(3) Anderson, T. F., 1949, The reactions of bacterial viruses with their host
Cells, Bot. Rev., 15, 464.

(3) Anderson, T. F., 1951, Tr. New York Acad. So., 13, 130.

(3) Anderson, T. F., and Doermann, A. H., 1952, J. Gen. Physiol., 35, 657.

(4) Satchwell SC, Drew HR & Travers AA (1986) chromosome &DNA. J Mol
Biol 191, 659–670.

(5) Brown, T. A. (2011). Introduction to genetics: A molecular approach. New

York: Garland Publishing.

 Crick, F. (1988). What Mad Pursuit: A Personal View of Scientific

Discovery. New York: Basic Books
 Wilkins, M. (2004). The Third Man of The Double Helix. Oxford University
Press.
 Judson, H. F. (1995). The eighth day of creation: Makers of the revolution
in biology. United Kingdom: Penguin Books.
 Gribbin, J. (1985). In Search of the Double Helix: Quantum Physics and
Life. United States of America: McGraw-Hill Book Company.
 Rsc.org,. "The DNA Story". N.p., 2016. Web. 25 Jan. 2016.

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 Watson, James, and Francis Crick. (1953). Molecular Structure Of Nucleic
Acids. Nature: n. pag. Print.
 Jaenisch, Rudolf, and Arnold Levine. (1971) DNA Replication In SV40-
Infected Cells. Virology 44.3: 480-493. Web.
 Wilmut, Ian. (2004) To Clone Or Not To Clone?. Endeavour 28.4: 135.
Web.
(6) Arnott S, Hutchinson F, Spencer M, Wilkins MH, Fuller W & Langridge R
(1966) X-ray diffraction studies of double helical ribonucleic acid. Nature
211, 227–230.
(7) Gartenberg MR & Crothers DM (1988) DNA sequence determinants
Nature 333, 44-54.

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