Bacteriophages

Dr.T.V.Rao MD

11/20/12 Dr.T.V.Rao MD's Undergraduate Series 1

 What are Bacteriophages

Viruses that attack bacteria were

observed by Twort and d'Herelle in
1915 and 1917. They observed that
broth cultures of certain intestinal
bacteria could be dissolved by addition
of a bacteria-free filtrate obtained from
sewage
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 Bacteriophages under Electron
Microscope

11/20/12 Dr.T.V.Rao MD's Undergraduate Series 4

 Bacteriophage (Phage)

Definition - Obligate intracellular parasites
that multiply inside bacteria by making use of
some or all of the host biosynthetic
machinery

Significance
− Models for animal cell viruses
− Gene transfer in bacteria
− Medical applications

Identification of bacteria - phage typing

Treatment and prophylaxsis???
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 Bacteriophages:
Definition & History

Bacteriophages are
viruses that can infect
and destroy bacteria.

They have been
referred to as bacterial
parasites, with each
phage type depending
on a single strain of
bacteria to act as host.
11/20/12 Dr.T.V.Rao MD's Undergraduate Series 6
 BACTRIOPHAGES

Like most viruses, bacteriophages

typically carry only the genetic
information needed for replication of
their nucleic acid and synthesis of their
protein coats.. They require precursors,
energy generation and ribosomes
supplied by their bacterial host cell.
11/20/12 Dr.T.V.Rao MD's Undergraduate Series 7
 Bacteriophages:
Classification
 At present, over
5000
bacteriophages
have been studied
by electron
microscopy and
can be divided
into 13 virus
families.
11/20/12 Dr.T.V.Rao MD's Undergraduate Series 8
 Bacteriophage
 Bacteriophages
make up a
diverse group of
viruses, some of
which have
complex
structures,
including double-
stranded DNA.
11/20/12 Dr.T.V.Rao MD's Undergraduate Series 9
 Bacteriophage

11/20/12 Dr.T.V.Rao MD's Undergraduate Series 10

 Bacteriophage
 Also known simply as a
phage; a virus that
attacks and infects
bacteria. The infection
may or may not lead to
the death of the
bacterium, depending on
the phage and
sometimes on
conditions. Each
bacteriophage is specific
to one form of bacteria.

11/20/12 Dr.T.V.Rao MD's Undergraduate Series 11

 Composition and
Structure

Composition
− Nucleic acid Head/Capsid

Genome
size

Modified
bases Contractile Tail
Sheath
• Protein
−Structure (T )
4

Size Tail Fibers

– 
Protection
Head or
–
Infection Base Plate
capsid
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–
Tail
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 Phage entering a bacterial cell

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Bacteriophage showing Lytic and
lysogenic cycle

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 Bacteriophages:
Virulence Factors Carried On Phage

Temperate phage can go through one of two
life cycles upon entering a host cell.
1) Lytic:
Is when growth results in lysis of the host and release
of progeny phage.
2) Lysogenic:
Is when growth results in integration of the phage DNA
into the host chromosome or stable replication as a
plasmid.
Most of the gene products of the lysogenic phage
remains dormant until it is induced to enter the lytic
cycle.
11/20/12 Dr.T.V.Rao MD's Undergraduate Series 17
 Bacteriophages:
Lysogenic Conversion

Some lysogenic phage carry genes that
can enhance the virulence of the
bacterial host.
 For example, some phage carry genes that
encode toxins.

These genes, once integrated into the
bacterial chromosome, can cause the
once harmless bacteria to release
potent toxins that can cause disease.
11/20/12 Dr.T.V.Rao MD's Undergraduate Series 18
 Bacteriophages
•
Used for cloning foreign
genes among other
applications
•
Proteins and peptides are
fused to the
Capsid(surface) of the
phage
•
The combination of the
phage and peptide is
known as a Fusion Protein

11/20/12 Dr.T.V.Rao MD's Undergraduate Series 19

 Bacteriophages
•
Used for cloning foreign
genes among other
applications
•
Proteins and peptides are
fused to the
Capsid(surface) of the
phage
•
The combination of the
phage and peptide is
known as a Fusion Protein

11/20/12 Dr.T.V.Rao MD's Undergraduate Series 20

 Bacteriophages
•
Once these Phages are isolated and
recovered they can be used to infect
bacteria which will create a particle similar
to a monoclonal antibody

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 Lytic and Lysogenic cycle

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 Bacteriophages:
Lysogenic Conversion
Examples of Virulence Factors Carried by Phage
Gene
Bacterium Phage Phenotype
Product
Vibrio cholerae CTX phage cholerae toxin cholera
lambda hemorrhagic
Escherichia coli shigalike toxin
phage diarrhea
clostridial botulinum botulism (food
Clostridium botulinum
phages toxin poisoning)
Corynebacterium corynephage diphtheria
diphtheria
diphtheriae beta toxin
Streptococcus erythrogenic
T12 scarlet fever
pyogenes toxins

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 Lysogenic conversion
In some interactions between lysogenic
phages and bacteria, lysogenic conversion
may occur. It is when a temperate phage
induces a change in the phenotype of the
bacteria infected that is not part of a usual
phage cycle. Changes can often involve the
external membrane of the cell by making it
impervious to other phages or even by
increasing the pathogenic capability of the
bacteria for a host.
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 Assay for Lytic Phage
Phage

•

Plaque assay
Plaque assay

 Method
Method
–
Bacteria
Plaque
–
forming unit +

Plaque
(pfu) forming Phage

unit (pfu) infectious

Measures
–

 particles
Measures
infectious
particles
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 Lytic
vs
Lysogenic
Cycle

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 Transduction

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Transduction

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 Bacterial Bacterial Capsid DNA
cell wall chromosome
Capsid

Sheath
Tail fiber
1 Attachment: Tail
Base plate
Phage
Pin
attaches to
host cell. Cell wall
Plasma membrane

2 Penetration:
Phage pnetrates
host cell and
injects its DNA. Sheath contracted

Tail core

3 Merozoites released
into bloodsteam
from liver may infect
new red blood cells
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Figure 13.10.1
 Tail
DNA

4 Maturation:
Viral components
are assembled Capsid
into virions.

5 Release:
Host cell lyses
and new virions Tail fibers
are released.

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Figure 13.10.2
 Examples:
* Corynebacterium diphtheria produces the
toxin of diphtheria only when it is infected by the
phage β. In this case, the gene that codes for the
toxin is carried by the phage, not the bacteria.
* Vibrio cholerae is a non-toxic strain that can
become toxic, producing cholera toxin, when it is
infected with the phage CTXφ.
* Clostridium botulinum causes botulism.
* Streptococcus pyogenes causes scarlet fever.
* Shiga toxin
11/20/12 Dr.T.V.Rao*MD's
Tetanus
Undergraduate Series 33
 Medical Applications of Phages
“I strongly believe phage could become
an effective antibacterial tool” - Carl Merril,
Chief of the Laboratory of Biochemical Genetics,
National Institute of Mental Health, NIH.

“It might be another string on the bow,
such that when (conventional antibiotics)
fail, here’s something that has a chance
of working. But it’s not going to be a
panacea” - Joshua Lederberg, Sackler Foundation
Scholar at The Rockefeller University

11/20/12 Dr.T.V.Rao MD's Undergraduate Series 34

 
The Programme Created by Dr.T.V.Rao MD
for Undergraduate Learning in Developing
Countries

Email

doctortvrao@gmail.com

11/20/12 Dr.T.V.Rao MD's Undergraduate Series 35