Genetic devices

Lecture 1
By
Alaa Hussein younus

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 The Genetic Material
Two different kinds of genetic material exist: deoxyribonucleic acid (DNA) and
ribonucleic acid (RNA). Most organisms primarily employ DNA, but a few viruses have
RNA as their genetic material. The biological information contained in an organism is
encoded in its DNA or RNA sequence.
-Prokaryotic genetic material is organized in a simple circular structure that resides in
the cytoplasm.
-Eukaryotic genetic material is more complex and is divided into discrete units
called genes.
-Human genetic material is made up of two distinct components: the
nuclear genome and the mitochondrial genome. The mitochondrial genome is a circular
DNA molecule separate from the nuclear DNA. Although the mitochondrial genome is
very small, it codes for some very important proteins.

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DNA was first extracted from nuclei in 1870, and named ‘nuclein’ after their
source. DNA Chemical analysis determined that DNA was a weak acid rich in
phosphorous. DNA name provides a lot of information about DNA
(deoxyribose nucleic acid):
– it contains a sugar moiety (deoxyribose),
– it is weakly acidic
–is found in the nucleus.
• Because of its nuclear localization and subsequent identification as a
component of chromosomes, it was implicated as a carrier of genetic
information

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Bacterial transformation implicates DNA as the substance of
genes:
In 1928 – Frederick Griffith – experiments with smooth (S)(virulent strain Streptococcus
pneumoniae), and rough (R)( nonvirulent strain of Streptococcus pneumoniae).
– Bacterial transformation demonstrates transfer of genetic material as shown in the figure
(1):
Griffith observed that live S bacteria could kill mice injected with them.
• When he heat killed the S variants and mixed them with live R variants,
and then injected the mixture in the mice, they died.
• Griffith was able to isolate the bacteria from the dead mice, and found them to be of the S
variety.
• Thus the bacteria had been Transformed from the rough to the smooth version.
• The ability of a substance to change the genetic characteristics of an organism is known as
transformation.
• Scientists set out to isolate this ‘transforming principle’ since they were convinced it was
the carrier of the genetic information.
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Figure (1): Griffith experiment: transformation of bacteria.
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• 1944 – Oswald Avery, Colin MacLeod, and MacIyn McCarty -determined that DNA is the transformation Material

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• Hershey and Chase experiment

In 1952 – Alfred Hershey and Martha Chase provide convincing evidence that
DNA is genetic material. Waring blender experiment using T2 bacteriophage
and bacteria Radioactive labels 32P for DNA and 35S for Protein.
The experiment was performed by using bacteriophage, a type of virus that have
a very simple structure: an outer core and an inner component. The phage are
made up of equal parts of protein and DNA. It was known that the phage infect
by anchoring the outer shell to the cell surface and then deposit the inner
components to the cell, infecting it.
• Scientists were interested in finding out whether it was the protein component
or the DNA component that got deposited inside the infected cell. By
Structure of T2 phage
incorporating radiolabel either in the protein or the DNA of the infecting
Bacteriophage = Virus that attacks
phage, they determined that the DNA was indeed introduced into the infected bacteria and replicates by invading a
causing proliferation of new phage. living cell and using the cell’s
molecular machinery.
Bacteriophages are composed of
DNA & protein

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Hershey and Chase experiment

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DNA is polar: 5’ to 3’ Beta N-glycosidic bond connects the sugar to the nitrogen base. A
phosphodiester bond connects one nucleotide to the next.
The sugar phosphate backbone is identical in every DNA molecule.
In any DNA purine = pyrimidine.
-The arrangement of the nitrogen bases determines the genetic message.

DNA is a polymer of nucleotides (monomers).

Each nucleotide has 3 parts:
1) 5 carbon sugar (Deoxyribose)
2) Phosphate group (PO4)
3) Nitrogen Base

Nucleotide
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Structure of DNA
James D. Watson/Francis H. Crick 1953 proposed the Double Helix Model based on two
sources of information:

1. Base composition studies of Erwin Chargaff (Chargaff’s Rules)

• indicated double-stranded DNA consists of ~50% purines (A,G) and ~50%

pyrimidines (T, C)

• amount of A = amount of T and amount of G = amount of C

• %GC content varies from organism to organism

Examples: %A %T %G %C %GC

Homo sapiens 31.0 31.5 19.1 18.4 37.5

Zea mays 25.6 25.3 24.5 24.6 49.1
Drosophila 27.3 27.6 22.5 22.5 45.0
Aythya americana 25.8 25.8 24.2 24.2 48.4

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James D. Watson/Francis H. Crick 1953 proposed the Double Helix
Model based on two sources of information:

2. X-ray diffraction studies by Rosalind Franklin & Maurice Wilkins

Conclusion-DNA is a helical structure with

distinctive regularities, 0.34 nm & 3.4 nm.

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Double Helix Model of DNA: Six main features

1. Two polynucleotide chains wound in a right-handed (clockwise) double-helix.

2. Nucleotide chains are anti-parallel: 5’ → 3’

3’  5’

3. Sugar-phosphate backbones are on the outside of the double helix, and the bases are
oriented towards the central axis.

4. Complementary base pairs from opposite strands are bound together by weak
hydrogen bonds.

A pairs with T (2 H-bonds), and G pairs with C (3 H-bonds).

5’-TATTCCGA-3’
3’-ATAAGGCT-5’

5. Base pairs are 0.34 nm apart. One complete turn of the helix requires 3.4 nm (10
bases/turn).

6. Sugar-phosphate backbones are not equally-spaced, resulting in major and minor

grooves.

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•Complementary base pairing:
– three hydrogen bonds between C and G;
–two hydrogen bonds between A and T.
Chargaff’s Rules
• Ratios of nucleotides
• A=T and G=C

• A+T does not have to equal G+C

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Eukaryotic chromosome structure
The chromatin is a complex of DNA and chomosomal proteins.

Two major types of proteins:

1. Histones abundant, basic proteins with a positive charge

that bind to DNA. There are 5 main types: H1, H2A, H2B, H3, H4

~equal in mass to DNA evolutionarily conserved.

2. Non-histones all the other proteins associated with DNA

differ markedly in type and structure , the amounts vary widely.

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Packing of DNA into chromosomes:

1. Level 1 Winding of DNA around histones to create a

nucleosome structure.

2. Level 2 Nucleosomes connected by

strands of linker DNA like
beads on a string.

1. Level 3 Packaging of nucleosomes into

30-nm chromatin fiber.

1. Level 4 Formation of looped domains.

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The many different orders of
chromatin packing that give
rise to the highly condensed
metaphase chromosome

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Euchromatin & Heterochromatin
Staining of chromatin reveals two forms:
– Euchromatin: condenses and decondenses with the cell cycle. It is
actively transcribed, and lacks repetitive sequences. Euchromatin
accounts for most of the genome in active cells.
– Heterochromatin remains condensed throughout the cell cycle. It
replicates later than euchromatin. is transcriptionally inactive.

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There are two types of heterochromatin
– Constitutive heterochromatin
• Regions that are always heterochromatic
• Permanently inactive with regard to
transcription
• occurs at the same sites in both
homologous chromosomes
consists mostly of repetitive DNA
– (e.g., centromeres).

– Facultative heterochromatin
• Regions that can interconvert between
euchromatin and heterochromatin
• varies between cell types or
developmental stages, or even
between homologous
chromosomes. Example: Barr body
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