Bacteriophages

• Bacteriophage (bacteria-eater), as the name

suggests, are the viruses that infect and replicate
within bacteria.

• They are commonly called a phage.

• They contain DNA or RNA in their genome, which is

encapsulated in a protein coat.
T4 (Escherichia virus T4)
• Linear dsDNA,
• Infects E.coli.
• Nonenveloped, Icosahedral head, tail is hollow with complex contractile structure
and has tail fibres.
• It undergoes only lytic life cycle, no lysogenic cycle.

T2 (Enterobacteria phage T2)

• Linear dsDNA,
• Nonenveloped, icosahedral head and have a contractile tail.
• It undergoes a lytic life cycle.

λ (Coliphage, Escherichia virus lambda)

• Linear dsDNA,
• The virus particle is made up of head, tail and tail fibres.
• Nonenveloped and have a noncontractile long tail.
• It undergoes both lytic and lysogenic cycles.
• Commonly used as a vector in recombinant DNA technology.

M13 (Escherichia virus M13)

• Circular ssDNA
• Filamentous, nonenveloped.
• Infects E.coli. M13 plasmids are used in genetic engineering.
ΦX174 or phi x 174 (Escherichia virus ΦX174)
• Circular ssDNA
• Isometric and nonenveloped. Infects E.coli.
• It was the first DNA genome to be sequenced and also the first genome to
be assembled in vitro.
• It encodes for 11 proteins.

Φ6, Φ7, Φ8
• Linear dsRNA,
• They are enveloped with icosahedral or spherical geometry.
• They contain protein and lipid outer layers.
• They mostly infect Pseudomonas bacteria.
T4 bacteriophage
• Phage binds to the lipopolysaccharide.

•
T4Tipsadsorption and injection
of the tail fibers contact the cell first.

• The base plate rearranges creating a hole in the base plate.

• Outer sheath contracts and the internal tube goes through the outer
membrane, peptidoglycan, and periplasm and comes close to the
cytoplasmic membrane.

• The DNA is injected and crosses the cytoplasmic membrane in about 30

seconds.

• After a 4-minute lag during which two proteins encoded by this piece of
DNA are synthesized, the rest of the phage DNA enters the cytoplasm.

• Binding of these two phage proteins to the DNA is thought to pull the DNA
into the cytoplasm.
T4 Bacteriophage
adsorption and DNA
injection
Life cycles of Bacteriophage
There are two ways by which bacteriophages infect the
host bacterium.

Lytic Cycle (Virulent infection)

• They induce complete lysis of the bacterial cell,
which is known as a lytic life cycle.

• Examples include T2, T4, T6 (T-even phages), they

are also known as virulent phages.

• The bacterial cell is completely destroyed

immediately after replication of the viral genome.

• This type of infection is called virulent infection and

it is mostly used for phage therapy.
The lytic cycle has the following steps:
1.Adsorption- Anchoring of bacteriophage to the
bacterial cell wall with the help of tails fibres.

2.Penetration- The phage DNA gets injected into

bacteria.

3.Replication and synthesis- The bacterial DNA is

disrupted and the viral genome takes charge of
bacterial machinery. It starts making proteins
required for replication and other structural proteins.

4.Assembly- Phage components are assembled into

new viral particles.

5.Lysis and release- Bacterial cells are lysed and new

viral particles are liberated to infect other cells.
Lysogenic Cycle (Temperate infection)

a. Attachment and Penetration

• Bacteriophages that undergo lysogeny are known as temperate phages.

• The viral DNA gets integrated into the host genome and replicates along
with the bacterial genome.

• The integrated viral genome is known as a prophage.

• It is relatively harmless and continues to remain in the position until the

lytic cycle is triggered.

• It may be spontaneous or due to certain external conditions such as

radiation exposure.

• Then the prophage becomes active and a lytic cycle initiates resulting in
the lysis of the cell wall.
• b. Replication
• After penetration, the phage DNA gets integrated into bacterial
DNA and gets replicated along with the bacterial genome.

• As the viral genome is inserted into the bacterial genome and

bacteria continue to perform the normal activities, the viral
genome gets transferred to the progenies as well.

• Bacterial cells containing a prophage are called lysogenic cells.

• Examples of lysogenic phage include lambda (λ) phage.

• Due to the ability to insert their genome specifically and

replicate, they are used in genetic recombination.
 Characteristics Lytic Cycle Lysogenic Cycle
The lytic cycle is a type of
Lysogenic is another type of
lifecycle of
lifecycle of bacteriophages
bacteriophages where the
which is defined by the
Definition viral DNA remains as a
incorporation of the
free-floating molecule
bacteriophage genome into
and replicates separately
the host genome.
from the bacterial DNA.

The lytic cycle is also The lysogenic cycle is also

Also called called the infective cycle called a temperate cycle or
or virulent cycle. non-virulent cycle.

In the lytic cycle, the viral In the lysogenic cycle, the

Viral DNA DNA remains in the viral DNA is incorporated
cytoplasm of the host cell. into the host chromosome.

No prophage is present in The lysogenic cycle consists

Prophage
the lytic cycle. of a prophage stage.

Host DNA is destroyed by

The host DNA is not affected
Host DNA various proteins encoded
by the viral DNA.
by the viral DNA.
 Characteristics Lytic Cycle Lysogenic Cycle

The viral DNA

Viral DNA replication
replication occurs
Viral replication occurs along with the
separately from the
host DNA replication.
host DNA replication.
The productivity of The productivity of viral
The productivity
viral DNA and viral DNA and viral proteins is
of viral DNA
proteins is high. low.
Host cellular
Host cellular mechanism is Host cellular mechanisms
mechanism completely hijacked by remain unaffected.
the viral DNA.
The lytic cycle is
immediate and is The lysogenic cycle takes
Duration
completed within a a longer period of time.
short period of time.
The lytic cycle cannot The lysogenic cycle can
Transition transition into a transition into the lytic
lysogenic cycle. cycle.
 Characteristics Lytic Cycle Lysogenic Cycle
As the cycle is an
infective cycle, The cycle is a non-infective
Infection symptoms of viral cycle that doesn’t result in
infections can be symptoms.
observed.
The viral DNA cannot be
The viral DNA can be
transferred from the
transferred into the
Transfer host cell to the daughter
daughter cell during the
cell during the lytic
lysogenic cycle.
cycle.

The lytic cycle doesn’t

The lysogenic cycle allows
Genetic allow genetic
the genetic recombination
recombination recombination of the
of the host chromosome.
host chromosome.

The lysogenic cycle doesn’t

The lytic cycle ends with
Lysis of host cell result in the lysis of the
the lysis of the host cell.
host cell.
Importance of Bacteriophage

• Bacteriophages are used for various purposes. They are widely

used in medical and research.
• Phage therapy- They are used as antibiotics against bacteria
due to the same mode of action.
• They are used in the food industry to kill bacteria in meat or
cheese products.
• Bacteriophages are used for diagnostic purposes.
• They act as a model in research and studies.
• They are used as a cloning vector in genetic recombination
technique
plaque assay.

The number of viruses in a sample can be

determined by direct count using electron microscopy
or by determining the number of infectious virus
particles using a plaque assay.
• Viruses are obligate intracellular parasites. Therefore, they require a host cell
in which to grow.

• Some viruses lyse the cells in which they have replicated while others appear
to cause little cell damage.

• A plaque assay can be used to enumerate viruses that lyse their host cells.

• In a plaque assay the host cells and virus are incubated together for a short
time to allow the virus to attach to and enter the host cell.

• Then the mixture in plated within a semi-solid agar.

• This semi-solid agar is poured onto a "bottom agar" that serves to supply
adequate nutrients for the host cell.

• At the end of one cycle of virus replication a cell infected with a single virus
particle will lyse, releasing hundreds of new viruses.
• In the semi-solid medium these newly released viruses can only infect
neighboring cells; after a second cycle of replication these neighboring cells will
be lysed.

• Those cells that escape infection will continue to grow. After 24 to 48 hours
plates that were not infected with a virus will contain a confluent layer of cells
(they will resemble the TNTC plates of the preceding exercise).

• Those plates that were infected with several hundred viruses may actually
appear clear- the viruses will have infected and lysed all of the host cells.

• Those plates that contain an intermediate number of viruses will have plaques,
clear or partially clear circular areas in an otherwise turbid background of cellular
growth.

• Each plaque represents the result of one infectious virus, called a plaque
forming unit, or PFU.

• Many animal and bacterial viruses can be enumerated using a plaque assay.
Aphids
icosahedral (20 sided) virus. This is because of the icosahedral shape of the
capsid that surrounds the viral genome. This shape gives the virus 20
symmetrical triangular face
Transmission

Cauliflower mosaic virus (CaMV) is transmitted by aphids

using the non-circulative transmission mode:

when the insects feed on infected leaves, virus particles

from infected cells attach rapidly to their stylets and are
transmitted to a new host when the aphids change plants.
Hepatitis B is an infection of the liver caused by the
hepatitis B virus.

The infection can be acute (short and severe) or

chronic (long term).

Hepatitis B can cause a chronic infection and puts

people at high risk of death from cirrhosis and liver
cancer.
Structure
Transmission