Bacteriophage

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

MS2
For lecture only

T2

Fd, M13
BC Yang

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) and Felix d'Herelle (1917)
independently reported isolating filterable entities capable
of destroying bacterial cultures and of producing small
cleared areas on bacterial lawns.
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).

For lecture only

BC Yang

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

For lecture only

BC Yang

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 circular
area of cell lysis in a turbid lawn
of cells.
Dynamic process

For lecture only

gal+

gal-

BC Yang

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. Delbrck (1939). The Growth of

Bacteriophage. J. Gen. Physiol. 22:365-384.

For lecture only

BC Yang

Lytic cycle of phage

3
1

For lecture only

BC Yang

Kinetics of phage infection

0 min. Attachment of T2 to a susceptible E. coli cell

1 min. Inject DNA into cell
1-7 min. Transcribe and translate early genes

7-15 min. Replication of phage DNA

10-20 min. Translation of phage late proteins (structural)

block bacterial DNA synthesis and degrade host chromosomal DNA

block transcription of host mRNAs
block translation of host proteins
small amounts of early proteins produced (catalytic functions)
transcription from single phage genome

transcribed from new phage DNA (many copies of template)

need large amounts of these proteins to build new virions

18-25 min. Assembly of new phage particles (end of eclipse

period)
25 min. Lysis of host cell and release of progeny (end of
latent period)

For lecture only

BC Yang

Infection processes

For lecture only

1.
2.

Attachment of virion to cell

3.

Early viral proteins synthesized (required

for genome replication)

4.
5.
6.
7.

Genome replication

Entry of viral nucleic acid into host cell (with

or without other virion components)

Late proteins synthesized (capsid proteins)

Assembly of progeny virions
Release of infectious progeny virions

BC Yang

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 Q)

DNA is the major material

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

For lecture only

BC Yang

Developmental gene expression

assay by protein synthesis

Early, in
5 min

For lecture only

Middle, in
10 min

Late
In 25 min
BC Yang

Host gene shut-off

Altering RNA
polymerase activity

Change the translation

apparatus (translation
of the MS2 phage RNA
with ribosome of T4infected cells reduced
by 88%)

Degradation of host
DNA

For lecture only

XP10

BC Yang

Assembly of phage
Can it happen automatically?

For lecture only

BC Yang

Lysogenic cycle
Lysogenic Cycle: Lambda as an example

For lecture only

lambda integrase and lambda repressor cI

synthesized due to activation of the
transcription of their genes by cII.
cI repressor turns off phage transcription
integrase catalyzes integration of lambda
DNA into bacterial chromosome via short
sites of homology (site-specific
recombination) ---- prophage
BC Yang

Return to be a killer

Prophage:

Prophage can be excised when host response

system to potentially lethal situations:

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.

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.

For lecture only

BC Yang

Application of phages

Model system of
molecular biology
Cloning and
expression
Phage display system
Phage typing
Phage therapy:

phage as natural, selfreplicating, selflimiting antibiotics.

For lecture only

BC Yang