Bacteriophage

-Lambda

Lambda Phage is a temperate phage

infect E.coli cells.
Structure of the virus particle consist of
Head
( icosahedral in shape ) and Long
flexible tail.
Head proteins encloses the 48.5kbp
linear double stranded DNA molecule.

 Binds to specific receptors on the outer

membrane of E.coli
Injects its linear genome through the
phage tail into the cell.
Once the linear double stranded DNA
entered in to the host cell, it can either
integrate into host chromosomal DNA and
becomes a noninfectious prophage , or
dont integrate but replicate their DNA
molecule and cause lysis of the host cell.

Linear and Circular forms of lambda DNA

Both the ends of linear ds DNA

molecule are flanked by single

stranded DNA of 12 nucleotides .
These single stranded DNA
molecules are complementary in
sequence
- referred as sticky ends or
cohesive ends are also called cos

cos site have two distinct roles during

infection cycle
1. Allow the DNA molecule to circularize
once entered into the host cell. ( This is
necessary for insertion into bacterial
genome)
2.

After

excision

of

prophage,

large

numbers of new lambda DNA molecules

are

produced

by

rolling

circle

Cos sites now act as recognition

sequences for endonuclease - which

cleaves catenane at cos sites.
- This results in the formation of
individual lambda genomes.

Endonuclease is a gene A
product of Lambda DNA

Which
creates
single
stranded sticky ends in lambda
DNA
also involved in packaging of
Lambda
genome
into
the
phage head in combination
with other proteins.

Gene Organization in the Lambda D

Genetic map of lambda phage shows

that the functionally related genes are

clustered together on the genome
Except gene N and Q
Map shows that genes coding for tail
and head proteins are grouped together
on left

 Central region responsible for

integration of lambda DNA in to host
genome
Most of the genes in this region
are non essential for phage growth
(Hence these genes are deleted or
replaced without impairing the
infectious growth cycle)

Life cycle of
Bacteriophage
Lambda (Lytic
and
Lysogenic)

Promoters and control circuits

In lytic cycle, lambda transcription occurs in three
stages:
1. Early gene products
establish the lytic
cycles
2. Middle gene products - Replication
3. Late gene products
- Assembly of new
virus
particles and lysis of
the cell
Lambda DNA uses host RNA polymerase for
the synthesis of proteins to undergo lysogenic
cycle

Early transcription starts from

promoters PL and PR
Fate of the viral genome, to undergo
lytic or lysogenic cycle is decided by the
cro and cI gene products
Between PL and PR repressor gene cI is
present and which inhibit the transcription
of PL and PR and allow the phage to
undergo lysogenic cycle

Transcription inhibition of PL and PR

occurs when the cI protein
binds to
operator sequences of early genes .

So the phage undergo lysogenic cycle.

If the cro proteins are able to occupy
the preferred regions of the operator
first, then stimulates its further synthesis
and blocks synthesis of the other protein.
Hence the lambda DNA undergo lytic
cycle.

Early transcripts from PL and PR

stops at tL and tR1

Site tR2 stops any transcripts that

escape beyond tR1
Shift from Early to middle stage
transcription is directed by N gene
product expressed from PL.

PL and PR transcripts extended to genes

red, O and P which are important for
middle stage.

When the enough cro gene products is

accumulated, it prevents the transcription
from PL and PR
Then gene Q products expressed from
PR directs middle to late shift .

Gene Q products is another antiterminator like gene N product.

So transcription of Late genes across
the cos region takes place.
results in many phage
particles .
Both the gene N and gene Q are
positive regulators essential for phage
growth and lysis.