MOLECULAR

GENETICS
THE CHEMICAL NATURE OF THE GENE

© 2016 Paul Billiet ODWS

What does a gene do?
• The gene must be able to hold
information and decode it (translate it) into
an organism as it grows and develops

• It must be able to copy itself so that it can

be passed on to future generations

© 2016 Paul Billiet ODWS

What does a gene look like?
• It must be a big molecule
to hold the large amount of information
required to build an organism

• It must be a complex molecule

to provide the necessary variation to code
the instructions that control growth and
development
© 2016 Paul Billiet ODWS
Four classes of molecules which could
form genes
Biological Elements Building Blocks
macromolecules
Polysaccharides CHO Monosaccharides
(carbohydrates)
Lipids CHO Fatty acids (and 
(Fats, oils and waxes) glycerol)
Polypeptides CHONS Amino acids
(proteins)
Polynucleotides CHONP Nucleotides
(Nucleic acids)
© 2016 Paul Billiet ODWS
Griffiths (1928)

Tried to determine
what genetic material
was made of.

profiles.nlm.nih.gov/CC/A/A/B/N/_/ccaabn_.jpg 
© 2016 Paul Billiet ODWS
 Griffiths’ Experiment
Pneumococcus bacteria on mice

2 STRAINS

S-type R-type
Smooth colonies Rough colonies
Virulent Avirulent

Innoculate into mice Innoculate into mice

Dead from Not killed

pneumonia
© 2016 Paul Billiet ODWS
Griffiths’ Experiment
EXPERIMENT CONTROL CONTROL
Live R­type  Live R­type only Heat­killed S­type 
(harmless) only
+
Heat­killed S­type
Mice died from  No mice died No mice died
pneumonia
Further test: 
Cultured lung fluid
Live S­type found

© 2016 Paul Billiet ODWS

Conclusion
• Transformation of R-type to S-type
• Transformation was brought about by some
heat stable compound present in the dead
S-type cells

Called the TRANSFORMING PRINCIPLE

© 2016 Paul Billiet ODWS

Avery, MacCleod & McCarthy (1944)

Tried purifying the

transforming principle
to change R-type
Pneumococcus to S-
type

© 2016 Paul Billiet ODWS

Results
The compound that had the most effect was:
• Colourless, viscous and heat stable
• It contains phosphorus
• It was not affected by trypsin (a protease)
or amylase.
• It was not inhibited by RNAase
• It was inhibited by DNAase

Conclusion
The transforming principle is a DNA
© 2016 Paul Billiet ODWS
 Live R­type
+
Experiment DNA extracted and purified 
from S­type bacteria

Mice died from 
pneumonia

Live S­type bacteria cultured 
from the lung fluid

These S­type bacteria remained 
virulent for generation after 
© 2016 Paul Billiet ODWS generation
Conclusion
DNA is the transforming principle and it is
hereditary material
Criticism
The DNA was not totally pure
It was contaminated by a small amount of
protein
This protein could be the real transforming
principle
BUT
Purer extracts of DNA were better at
transforming the bacteria types.
© 2016 Paul Billiet ODWS
Hershey & Chase (1952)

To determine the nature of hereditary

material used by viruses

© 2016 Paul Billiet ODWS

Virus
A virulant sub-
microscopic agent
which will pass
through filters that
normally stop bacteria.

Ebola virus
© 2016 Paul Billiet ODWS
Virus

• Luria (1955) “bits of heredity in search of

chromosomes”

• Lwoff (1966) “viruses are viruses because they

are viruses”.

© 2016 Paul Billiet ODWS

Virus
Three properties of all viruses
1.Infectivity
2.Absence of biochemical apparatus (no
metabolism). A virus is not alive
3.Possess only one type of nucleic acid
(DNA or RNA).

© 2016 Paul Billiet ODWS

Types of virus
• Positive RNA viruses: TMV & Enterovirus (polio)
RNA used like mRNA directly by the host cell for
translation.
• Negative RNA viruses: Ebolavirus, Orthomyxoviridae
(influenza)
Viral RNA is transcribed into positive RNA before it is
translated. The RNA polymerase that does this is supplied
by the virus.
• Retrovirus: HIV
Inject RNA strand plus reverse transcriptase. A DNA
copy is made and used for the expression of viral genes.
• Bacteriophages: DNA viruses.

© 2016 Paul Billiet ODWS

Virus structure
• Composition: Protein
and nucleic acid
• Organisation : Protein
coat covering a nucleic
acid core
• A virus does not
posses a cellular
structure
• Crystallisable.
© 2016 Paul Billiet ODWS
© 2016 Paul Billiet ODWS
© 2016 Paul Billiet ODWS
T2 Phage particle
E. coli bacterium before infection

© 2016 Paul Billiet ODWS

4 min

© 2016 Paul Billiet ODWS

10 min

© 2016 Paul Billiet ODWS

12 min

© 2016 Paul Billiet ODWS

30 min

© 2016 Paul Billiet ODWS

 Virus “life” Lysis

cycle

Packaging

Infection
Protein 
synthesis

Replication
© 2016 Paul Billiet ODWS
The Herschey & Chase
Experiment
• Proteins: CHONS
labeled with 35S
• Nucleic acids
CHONP labeled with
32P

Phage particle

© 2016 Paul Billiet ODWS

Milk shakes at Cold Spring Harbour
The Waring Blender

© 2016 Paul Billiet ODWS

The Herschey & Chase
Experiment
• Two types of T2 phages prepared:
S labeled (proteins labeled)
35

32P labeled (nucleic acids labeled)

• Infect fresh E. coli host cells with each type

• Centrifuge to remove the surplus phages
• Agitate in the blender to remove phages coats from
host cells
• Centrifuge again and collect:
(a) phage coats
(b) bacterial cells.
© 2016 Paul Billiet ODWS
Results

Destination of the
Centrifuge radioactivity / %
fraction P labeled
32
S labeled
35

phages phages
Bacterial host 80 20
cells
Phages 20 80
© 2016 Paul Billiet ODWS
Observation

Most of the 32P (i.e. DNA) is injected into the

host but not much 35S (i.e. protein).

© 2016 Paul Billiet ODWS

Error margin
• The 20% P in the phage coat fraction
35

phages that had not been removed by the

first centrifugation plus attached phages
that had not yet injected their DNA

• The 20% S in the bacterial cell fraction

35

could be due to phage coats which had not

been successfully knocked off by the
blender.
© 2016 Paul Billiet ODWS
 Going further
• Of the P which got into the host cells, half of it
32

was found in phages produced from infected cells

• Less than 1% of the 35S appeared in the next
generation viruses.

© 2016 Paul Billiet ODWS Phage infecting a bacterium