BACHTERIOPHAGE

Phage T4 infected bacterium E.coli

Phage T4 Phage tail

Fedik A. Rantam, DVM., Dr., Prof.

Lab Virology and Immunology, Dep of
Microbiology, FVM-UA

08/14/2020 Rantam, 2018

 BACTERIOPHAGE POSITION
MICROBIOLOGY

BACTERIOLOGY MICOLOGY VIROLOGY IMMUNOLOGY

VIRUS

HUMAN, ANIMAL, BACTERIUM

PLANTES

BACTERIOPHAGE ?
 BACTERIOPHAGE ?

 Virus, who is infected bacterium

 Their properties are specific (receptor)
 They have taxonomy and morphology
 They can replicate in bacterium
 Application of bacteriophage

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 Bacteriophage
 Prokaryotes as host
 Subcellular structure without metabolic machinery
 Double stranded DNA, single stranded DNA, RNA
 Virulent phage vs. template phage

MS2 T2 Fd, M13

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 CLASIFICATION OF BACTERIOPHAGE

Morphology :
 Icosahedral: tail contactile, T5 and, non tail, non
contractile
 Phage filament: phage enveloped, Pleomorphic,
Isometric head, elongated head
 Nucleic acid properties : DNA (deoxyribonucleat acid),
RNA (Ribonucleat acid)

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 Familly and Genus
Bacteriophage :
1. DNA (dsDNA):
Myoviridae,
Siphoviridae,
Podoviridae,
Corticoviridae,
Plasmaviridae,
Fuselloviridae,
Tectiviridae, Rudiviridae,
Lipothrixviridae
2. DNA (ssDNA):
Microviridae, Inoviridae

3. RNA : dsRNA Cystoviridae

4. ssRNA : Leviviridae
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 CENTRAL DOGMA

AMINO
RNA/DNA PEPTIDE
ACID

POLIPEPTI
PROTEIN EKSPRESI
DE

EFEK

Varian
Plaque Lysis
baru

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 History of bacteriophage Definition of bacteriophage

Bacteria E. coli
Application of Startegy of infection
bacteriophage

Phage T4 Growing curve of

Return to be killer Bacteria E. coli were infected bacteriophage
phage after 30 men

Lytic strategy

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 3. Historical context
 A century ago, Hankin (1896) reported that the waters of the
Ganges and Jumna rivers in India had marked antibacterial
action (against Vibrio cholerae, restrict epidemic) which could
pass through a very fine porcelain filter; this activity was
destroyed by boiling.
 Edward Twort (1915, staphilococcus) and Felix d'Herelle (1917,
coccobacillus) independently reported isolating filterable
entities capable of destroying bacterial cultures and of
producing small cleared areas on bacterial lawns. Maxdelbruck
and Emory Ellis (1939, suitable medium for phage culture)
 It was F d'Herelle, a Canadian working at the Pasteur Institute
in Paris, who gave them the name "bacteriophages"-- using the
suffix phage (1922).
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 plaque
 Plaques are clear zones
formed in a lawn of cells due
to lysis by phage.
 At a low multiplicity of
infection (MOI) a cell is
infected with a single phage
and lysed, releasing progeny
phage which can diffuse to
neighboring cells and infect
them, lysing these cells then
infecting the neighboring
cells and lysing them, etc,
 It ultimately results in a gal+
circular area of cell lysis in a
turbid lawn of cells.
 Dynamic process gal-

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 Glossary
 pfu: plaque forming unit
 Title: define pfu in a phage suspension
 moi: multiplicity of infection, the ration of phage
particles to bacteria
 eop: efficiency of plating, the ration of the plaque titer
to the number of phage particles
 Prophage: state of phage co-existing with host
 Lysogenic bacteria: term of bacteria carrying prophage
 Phage conversion: phenotype change in lysogenic
bacteria

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 Infection processes

1. Attachment of virion to cell

2. Entry of viral nucleic acid into host cell
(with or without other virion
components)
3. Early viral proteins synthesized (required
for genome replication)
4. Genome replication
5. Late proteins synthesized (capsid proteins)
6. Assembly of progeny virions
7. Release of infectious progeny virions

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 Bacteriophage Growth Curve
Latent Period Rise Period Latent phase: fast growth,
without virion
Phage Count

Rise phase: hight

population phage in the
cell, Infective phage:
release phage from the
cell

Time (Minutes)
Receptor
specific :
Lypopolysacarid
dan protein
(T4), Teichoic
T4 Phage Adsorption and DNA acid, Flagella
dan filly-filly
Injection
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 One step growth

Demonstrate an eclipse period during which the DNA began

replicating and there were no free phage in the cell, a period
of accumulation of intracellular phage, and a lysis process
which released the phage to go in search of new hosts.

Ellis, E. L. and M. Delbrück (1939). The Growth of Bacteriophage.

J. Gen. Physiol. 22:365-384. 
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 Lytic cycle of phage

E.coli

6 7
5
3
1 4

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 Return to be a killer

 Prophage:
 Bacterium is now immune to infection by another phage,
because repressor continuously produced ----- new phage DNA
can be injected into cell and is circularized but is not transcribed
or replicated.
 Prophage can be excised when host response
system to potentially lethal situations:
 if host DNA damaged
 one reaction by host cell is to activate a protease
 protease also cleaves repressor
 Phage DNA now transcibed including a gene for an enzyme that
cuts prophage DNA from bacterial chromosome
 Lytic cycle can start.

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 Adsorption and DNA injection

 A random collision,
protein/protein interaction
 Affected by Ca++, Mg++, or
tryptonphanetc.
 Receptor specific (outer
membrane protein lamB for
lambda; sex pili for Qb)

 DNA is the major material

entering bacterial
 Lysozme like activity, core
boring through the cell wall

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 Strategy of Infection (Adsorption phage on
Bacterium)
 Fiber Binding with receptor
 Fiber landed on outer membrane using electrostatic
 changed pH and ion-ion Mg2++ and Ca2++
 Changing of phage conformation
 Reformed of 24 ring to 12 ring
 DNA spread into the cell through interaction phage and
plasma membrane

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 Developmental gene expression
assay by protein synthesis

Early, in
5 min
Middle, in
Late
10 min
In 25 min

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 Application of phages

 Model system of
molecular biology
 Cloning and
expression
 Phage display system
 Phage typing
 Phage therapy:
 phage as natural, self-
replicating, self-
limiting antibiotics.

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 Developmental gene expression
assay by protein synthesis

Early, in
5 min
Middle, in
Late
10 min
In 25 min

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 Host gene shut-off

 Altering RNA
polymerase activity
 Change the translation
apparatus (translation
of the MS2 phage RNA
with ribosome of T4-
infected cells reduced
by 88%)
 Degradation of host
DNA XP10

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 Detection of Bacteriophage

 Plaque
 Lysed culture
 SDS – Page (electrophoresis)
 Polymerase Chain Reaction (PCR/ RT-PCR)
 Mutiplex PCR

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 Bac. Application

 Rapid multiplex detecvtion Endolysis using

bacteriophage for listeria spp
 Elimination of upper respiratory colonization
streptococci
 Endolysin EL 188 and outermembrane permeability
against Pseudomonas aeroginosa
 Enhance lytic activity and binding activity
 As vector for Treatmen antimycrobial resistance in
food

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 VIROLOGY AND IMMUNOLOGY
LAB, DEP.of MICROBIOLOGY,
VET.MED.FACULTY
UA- SURABAYA

Thank you/ Danke Schoen

08/14/2020 Rantam, 2017

 Assembly of phage
Can it happen automatically?

08/14/2020 Rantam, 2017

 Live cycles Bacteriophage
T4 : some menutes T4
injected its DNA.

bacteriophage T2: some phage attachment

bacterium and injected its DNA
 Assembly of BACTERIOPHAGE T4:
1. Late mRNA were produced after DNA
replication
2. Synthesis of 3 proteins : structural
protein for build virion and release
phage.
3. Protein synthesis for independent
assembly
4. Base plate were constructed by 15
gene, head were constructed by 10
protein with build tail together
5. Portal Protein : head assembly and
support DNA in and out from head
masuk dan keluar kepala

Assembly of Phage
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 SELF READING

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