Biomedical Engineering Reference
In-Depth Information
Gene 8 protein
F pilus of F
+
E.coli
Gene 3 protein
Stage 1
Absorption and
conversion of
single-stranded
DNA to double-
stranded
(replicative form,
RF) DNA
0-1 minute
post-infection
M13 filamentous phage particle
(870
×
6 nm). Single-stranded DNA
circle of 6407 nucleotides
5
(+)
2
RNA
pol
4
3
DNA
pol
III
Nicking by gene
2
protein
Stage 2
RF
→
RF rolling
circle replication
5
6
∼
20 minutes
post-infection
8
(+)
7
Cutting by
gene
2 p
rotein
Stage 3
Gene 5 protein
switches DNA
synthesis to
production of
single strands
Gene 5 protein
∼
20 minutes
post-infection
to indefinitely
(+)
Virus particles extruded from
infected cell without lysis
Fig. 4.16
Life cycle and DNA replication of phage M13.
transform a strain of
E. coli
with a deletion of the β-
galactosidase α-fragment and recombinant phage
detected by intragenic complementation on media
containing IPTG and Xgal (see Box 3.2 on p. 35).
One of the blue plaques was selected and the virus in
it designated M13 mp1.
Insertion of DNA fragments into the
lac
region of
M13 mp1 destroys its ability to form blue plaques,
