1

BACTERIOPHAGES

PHAGE T4
HEAD

4 CHROMOSOME: DOUBLE STRAND LINEAR DNA ~2 x 105 NUCLEOTIDE PAIRS ~1 x 108 MOLECULAR WEIGHT ~200 GENES

CAPSOMER

TAIL

CORE SHEATH
TAIL FIBER (6)

COLLAR
BASE PLATE

EXTERIOR SPIKES NOBEL HERSHEY

INJECTION PENETRATION
RECEPTOR PROTEIN

WALL - OUTER MEMBRANE CYTOPLASMIC MEMBRANE

CYTOPLASM

PHAGE PLAQUES
~107 HOST CELLS

TOP AGAR

AGAR PLATE

INCUBATE

CONFLUENT GROWTH

PHAGE PLAQUES
~107 HOST CELLS ~10 T4 AND ~107 HOST CELLS

TOP AGAR

TOP AGAR

AGAR PLATE

INCUBATE

CONFLUENT GROWTH

PLAQUES

PLAQUE FORMATION BY LYTIC (VIRULENT) PHAGE

FREE PHAGES 30 MIN

HOST CELLS INFECTED CELL

MANY CYCLES

PLAQUE APPEARS CLEARHOST CELLS DESTROYED PHAGE INVISIBLE

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GROWTH IN LIQUID CULTURE (OF HOST)

LOG TURBIDITY

~1 x 108 CELLS / ML

TIME

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GROWTH IN LIQUID CULTURE (OF HOST)

LOG TURBIDITY

~1 x 108 CELLS / ML

ADD 2-3 x 108 T4 / ML TIME

PHAGE TITER?

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REPRESSOR

PROPHAGE

BINARY FISSION

LYSOGENS; LYSOGENIC; PASSIVE REPLICATION OF PROPHAGE DURING BINARY FISSION OF HOST

TURBID PLAQUE FORMATION BY TEMPERATE PHAGE

FREE PHAGES 30 MIN LYSOGENS

22

HOST CELLS INFECTED CELL

MANY CYCLES

STAGES:

6
ADSORPTION / ATTACHMENT SPECIFICITY-RECEPTORS INJECTION / PENETRATION SHEATH CONTRACTS CHROMOSOME INJECTION SYNTHESIS OF COMPONENTS (REPLICATION)
TRANSCRIPTION TRANSLATION ENERGY PROVIDED BY HOST PRECURSORS RIBOSOMES

1. 2.

.3

PRODUCES: VIRAL mRNA VIRAL DNA (RNA) CHROMOSOME VIRAL STRUCTURAL PROTEINS ASSEMBLY / MATURATION DNA PACKAGED TAILS ADDED INTACT VIRUSES PRODUCED .4

LYSIS / RELEASE T4 LYSOZYME PEPTIDOGLYCAN HYDROLYSIS

SUMMARY: ONE PHAGE-INFECTED CELL PRODUCES ~102 PROGENY IN ONE GROWTH CYCLE BURST SIZE: AVERAGE NUMBER PROGENY / INFECTED CELL

.5

PHAGE LAMBDA - l - TEMPERATE LYTIC GROWTH OR LYSOGENY 48,502 BP 30 GENES

15

THE l CHROMOSOME COHESIVE SITE

l DNA INTEGRATION
ATTACHMENT SITE

18

HOST CHROMOSOME

GALACTOSE GAL= BIOTIN OPERON BIO =

REPRESSOR [REPRESSION]

INTEGRASE

PROPHAGE LYSOGEN, STABLE, LYSOGENY PASSIVELY REPLICATED

INTEGRATION DETAILS
5' 5' 3' 3'
INTEGRASE CUTS BOTH COMMON CORES

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HOST DNA l DNA

REJOINS l AND HOST DNA

l PROPHAGE

LYSOGENIC IMMUNITY
[l WILL NOT GROW ON A l LYSOGEN] PROPHAGE REPRESSOR l COMES FROM OUTSIDE AND INFECTS

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NO l REPLICATION

LYSIS / LYSOGENY DECISION

A. ESTABLISHING LYSOGENY AFTER INFECTION REQUIRES: cI cII, cIII, INT
CRO ANTI-cI REPRESSOR (REQUIRED FOR LYTIC GROWTH) B. MAINTAINING LYSOGENY DURING HOST GROWTH REQUIRES: cI ESTABLISHMENT HOST RNA POLYMERASE BINDS PR AND PL; A. INFECTION TRANSCRIBES EARLY GENES CRO, cII, cIII STIMULATES RNA POLYMERASE TO TRANSCRIBE FROM PRE PRODUCES cI AND FROM PI INTEGRASE [BINDS UNIQUE -35 REGIONS] [cIII PREVENTS cII DEGRADATION] B. cII -

BINDS OR3 - LIMITS TRANSCRIPTION FROM PRM BINDS OR2 OR1 OL3 OL2 OL1 - TURNS OFF EARLY FUNCTIONS BINDS OR1 > OR2 COOPERATIVELY INHIBITS LYTIC GENE EXPRESSION [CRO, cII, OP, etc] STIMULATES PRM

C. CRO D. cI -

MAINTENANCE cI STIMULATES ITS OWN TRANSCRIPTION AND INHIBITS ALL OTHERS

REGULATORY PROTEINS: cI - REPRESSOR (FACILITATES LYSOGENY) BINDS OR1 > OR2 > OR3 BINDS OL1 > OL2 > OL3 - ACTIVATOR OF PRE AND PI cII DEGRADATION PREVENTS cII cIII -

OPERATORS / PROMOTERS: OR/OL - THREE OPERATORS EACH PR AND PRM - OVERLAP OR PL - OVERLAPS OL PRE - PROMOTER FOR REPRESSION ESTABLISHMENT PRM - PROMOTER FOR REPRESSION MAINTENANCE

GROWTH) CRO - ANTI-cI REPRESSOR (FACILITATES BINDS OR3 > OR2 > OR1 BINDS OL3 > OL2 > OL1

RNA 1. l DNA PENETRATES, CIRCULARIZED, HOST FUNCTIONS POLYMERASE TRANSCRIBES EARLY

2. cII ACTIVATOR STIMULATES TRANSCRIPTION FROM PRE, PI

3. CRO BINDS OR3; PREVENTS PRM FROM FUNCTIONING

4. cI REPRESSOR BINDS OR1 OR2 ; SHUTS OFF EARLY GENES TURNS ON PRM OL1 OL2

[CRO / cI RATIO IS CRITICAL]