Chapter # 17

The Viruses: Viruses of Bacteria &

Archea

C H A P T E R 17 A scanning electron
Lecture # 18 micrograph of T-
Topics to be covered include
even
• Structure of bacteriophage
bacteriophages
• One-step growth exp
infecting E. coli.
The
phages are colored
blue
Bacteriophage families

There are four different types of genetic material in case of

bacteriophage.
1. dsRNA
2. ssRNA
3. dsDNA
4. ssDNA
So, on the basis of genetic material, here are different families.
• Viruses are acellular and non-living.
• They can be living if they get host.
• Also called even
bacteriophages.
• The upper portion is capsid,
that contain genetic material
(it can be DNA or RNA).With
the aid of collar it is attached
to lower part. The lower
circular plate is called base
plate that contain tail pins
and tail fibers.
• Bacteriophage can be isolated
from water sources, soil,
plants, animals, and sewage.
BACTERIOPHAGE LIFE CYCLE
ANIMATION
Bacteriophage isolation
 Reproduction of Double-Stranded DNA
Phages: The Lytic Cycle
• Lytic cycle
• T-even phages of E. coli
• Double stranded DNA bacteriophages with complex contractile
tails (family Myoviridae)
Note:
E.Coli is affected by T4 bacteriophage and cause its lysis.
 The One-Step Growth Experiment(used to visualize
all stages)

An experiment used to study the reproduction of lytic phages in which one

round of phage reproduction occurs and ends with the lysis of the host
bacterial population.
1) Mixing of E. coli is with bacteriophage particles followed by incubation
2) Dilution of culture mixture
3) Release of infective phage particles
4) Plaque count
Note:
This process ensure that each bacteriophage infect each bacteria.
Took E.coli from petri dish and add it in nutrient broth. Petri dish contain already prepared inoculum of 18
hours present in refrgirator. We do not directly use E.Coli from petri dish bcz we do not know on which stage
they are. Now incubate it. Dilute the culture mixture by the same media. After some time, phage particles will
release. Now phage count(we do colonies count CFU in case of bacteria).
 One-step growth curve experiment
Figure 17.2 The One-Step Growth Curve. In the
initial part of the latent period, the eclipse period, the
host cells do not contain any complete, infective
virions. During the remainder of the latent period, an
increasing number of infective virions are present,
but none are released. The latent period ends with
host cell lysis and rapid release of virions during the
rise period or burst. In this figure the blue line
represents the total number of complete virions. The
red line is the number of free viruses (the
unadsorbed virions plus those released from host
cells). When E. coli is infected with T2 phage at 37°C,
the growth plateau is reached in about 30 minutes
and the burst size is approximately 100 or more
virions per cell. The eclipse period is 11–12
minutes, and the latent period is around 21–22
minutes in length.
 The One-Step Growth Experiment
1. Latent period follows phage addition, there is no release of virions.
2. Rise period or burst, host cells rapidly lyse and release infective
phages.
Plateau is reached and no more viruses are liberated.
Burst size, the number of viruses produced per infected cell
The One-Step Growth Experiment
1. Latent period (shortest time) required for virus reproduction
and release.
2. Lysis of bacteria with chloroform.
3. Eclipse period because the virions were detectable before
infection but are now concealed or eclipsed.
The One-Step Growth Experiment

When the experiment is carried out with actively growing

cells in rich medium at 37°C, the growth curve plateau is
reached in approximately 30 minutes.
Bacteriophage life cycle (d)

1) Adsorption
2) Dilute culture
3) Samples
4) Assay
(a) (b)

(c)