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TEMPERATE

BACTERIOPHAGE

TEMPERATE BACTERIOPHAGE
TEMPERATE BACTERIOPHAGE

TEMPERATE BACTERIOPHAGE

Stable relationship with the host

Phage DNA replicates together with the host DNA

Late phase proteins are not synthesized

Lytic or lysogenic cycles

No degradation of the host DNA

Modification of the host RNA polymerase

Cells carrying the temperate phage = lysogen

Phage DNA + bacterial DNA = prophage

Lysogenic cycle lytic cycle – spontaneously or when the host metabolism is disturbed

LYSOGENIC CYCLE

1. Phage adsorption, injection of DNA

2. DNA circularization

3. Integration into the host DNA

4. Replication of host and phage DNA

5. Transcription of host and phage DNA

6. Induction by stress, chemicals

7. Excision of the phage DNA from the prophage

8. Degradation of host DNA, modification of the polymerase

9. Early and late phase transcription

10. Assembly of phage particle

Excision from the prophage DNA during induction into the lytic cycle requires excionase, integrase

Transcription is controlled by cro, cII proteins – determine whether lytic or lysogenic cycle is initiated

LYSOGENY

Circularization of the phage chromosome – cohesive ends

Cohesive Ends Linear Double Stranded O p e n e d C i r c

Cohesive Ends

Linear Double Stranded

Cohesive Ends Linear Double Stranded O p e n e d C i r c l

Opened Circle

Lygase

Cohesive Ends Linear Double Stranded O p e n e d C i r c l
Cohesive Ends Linear Double Stranded O p e n e d C i r c l

Closed Circle

Site specific recombination

Enzyme is coded by the phage

Suppression of phage genome expression repressor protein (cI)

Specific

Immunity to

super-infection (second phage)

LYSOGENY

gal bio gal bio gal bio
gal
bio
gal
bio
gal
bio

Map of Lambda Phage

Map of Lambda Phage

STOPPING LYSOGENY

Induction

Environment is not

suitable

Role of protease

recA Protein

Degradation of

repressor protein

cI

Excision Lytic growth

gal bio
gal
bio
gal bio
gal
bio
gal bio
gal
bio
gal bio gal bio gal bio gal bio
gal bio
gal
bio
gal bio gal bio gal bio gal bio

CHOICE BETWEEN LYTIC AND LYSOGENIC CYCLES

P L , P R : Promoter to start a competition between both cycles – synthesis of cro and N proteins

N Protein : regulator protein

N binds to t L , t R1 , t R2 - synthesizes cII, O, P and Q

cII: activator protein

O,P: protein to start λ DNA synthesis

Q: lytic cycle regulator protein

N binds to t L – synthesizes int, xis, cIII

int: integrase

xis: excisionase

cIII: protein to stablize cII

cIII, cII, O, P, Q: early phase genes

LYSOGENIC CYCLE

cII-cIII complex:

Binds to PRE – activates cI transcription (λ repressor)

cI: binds to P 1 – activates int transcription

λ repressor binds to O R , O L - prevents transcription from P R , P L – PREVENTS THE LYTIC CYCLE

cI is transcribed from P RM - synthesizes λ repressor

LYTIC CYCLE

Cro protein binds to O R , inhibits transcription from P RM – no synthesis of λ repressor

Transcription from P R produces O, P, Q proteins

O,P, catalyze λ DNA replication

Q: instructs transcription from P R' synthesize structural proteins and lysozyme

λ BACTERIOPHAGE

Host = E. coli

Temperate phage

Sticky cos ends

Host DNA ligase circularizes phage DNA

Lytic or lysogenic cycle

Genes are arranged according to function

Host RNA polymerase is modified

Replication by rolling circle or theta

Sequential transcription

Integrates into the host genome by recombinatian