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BACTERIOPHAGES
DEFINITION
• Bacteriophage, also called phage or
bacterial virus, group of viruses that
infect bacteria
• more than 10³¹ bacteriophages
HISTORY
• Frederick W. Twort in
Great Britain (1915) –
Staph col – degen.
• Félix d’Hérelle in France
(1917) – dysentry
• D’Hérelle coined the
term bacteriophage,
meaning “bacteria eater”
• (Phagein) - "to devour"
Félix d’Hérelle
Frederick W. Twort
INTRODUCTION
• Wide distribution
• Faeces, sewage, Natural
water
• Study – Virus host
interaction
• Transduction
• Phage conversion
• Phage typing – Id bacteria
CLASSIFICATION
• International Committee
on Taxonomy of Viruses
(ICTV) according to
morphology and nucleic
acid.
• Nineteen families
• 2 families have RNA
genomes
• 5 families – envelope
• 8 families with DNA
genomes - circular
genomes
• 9 (DNA) - linear
genomes
• Myoviridae
• Contractile tail
• Linear ds DNA
• T4 phage (T even
phage) – E. coli
MORPHOLOGY
• Tadpole
• Hexagonal Head
• Cylindrical tail
• Head –
– Tightly packed core of
ds DNA
– Surrounded by Protein
coat / Capsid
• Tail –
– hollow core
– Contractile sheath
surr
– Terminal base plate –
attached prongs /
fibres
• Spherical / filamentous
• ss DNA / RNA
LIFE CYCLE
LYTIC PHASE
• Virulent
• Intracellular
multiplication
• Bacterial host – lysis
• Release of progeny
virions
LYSOGENIC PHASE
• Temperate
• Phage DNA combines
with host s DNA
• Replicating together
• No harm to host
LYTIC CYCLE
• Adsorption
• Penetration
• Synthesis of phage components
• Assembly
• Maturation
• Release
ADSORPTION
• Attaches to bacterial surface to
area having complimentary
receptors to its base plate
• Rapid – minutes
• Location of bacterial Rs – cell
wall / Flagella / Pili
• Protoplasts – Not infected
( Transfection )
PENETRATION
• Lysozyme on tail makes hole on surface
• Hollow core pierces the cell wall
• Contractile tail sheath propels nucleic acid in bacteria
• Initiates synthesis of daughters
• Empty head and tail remain outside – GHOST
SYNTHESIS OF PHAGE COMPONENTS
• First synthesised – Early proteins
• Enzymes – manufacture of complex molecules
– phage specific
• Late proteins – Head and Tail
ASSEMBLY
• DNA of daughter phage – Packed into head
• Tail is added
MATURATION
• The previous process and conversion to
infective stage
RELEASE
• Lysis of bacterial cell
• Phage enzymes
• Bacteria becomes spherical during replication of
Phage
• Defective cell wall
• ECLIPSE PHASE – Entry to first intracellular
phage formation
LYSOGENIC CYCLE
• Symbiotic Relationship
• Enters the bacterial cell
• Integrated in bacterial chromosome
• Prophage
• Replicates synchronously with host
DNA
• Lysogeny
• Bacterial host metabolism -
UNAFFECTED
• Lysogenic / Phage conversion –
– Prophage confers new properties to bacteria
– ? - synthesis of new proteins
– Eg – toxin production in Diptheria by beta
phage
• During repeated multiplication – lysogenic phage
may be excised from host chromosome – initiates
– Lytic cycle – SPONTANEOUS INDUCTION
OF PROPHAGE
SPONTANEOUS INDUCTION OF PROPHAGE
• UV Rays
• Hydrogen peroxide
• Nitrogen Mustard
• Lysogenic bacteria – Resistant to reinfection –
Superinfection Immunity
TRANSDUCTION
• Carrier of genes between 2 bacteria
• 2 Types
– Restricted – genes common to bacteria
– Generalised – Any
• Plasmid mediated drug resistance - MRSA
PHAGE ASSAY
Zone of lysis after incubation – Plaques
Size, shape, nature – characteristic for different phages
PHAGE ASSAY
• Titrating the number of viable phages
• Purification of phages
PHAGE TYPING
Specificity of Phage bacterium interaction - Id and Typing of Bacteria
Adaption of Host – after serial passage in host becomes specific for
that host
APPLICATIONS
Intraspecies typing of bacteria
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Bacteriophages and their general characteritics

  • 2. DEFINITION • Bacteriophage, also called phage or bacterial virus, group of viruses that infect bacteria • more than 10³¹ bacteriophages
  • 3. HISTORY • Frederick W. Twort in Great Britain (1915) – Staph col – degen. • Félix d’Hérelle in France (1917) – dysentry • D’Hérelle coined the term bacteriophage, meaning “bacteria eater” • (Phagein) - "to devour" Félix d’Hérelle Frederick W. Twort
  • 4. INTRODUCTION • Wide distribution • Faeces, sewage, Natural water • Study – Virus host interaction • Transduction • Phage conversion • Phage typing – Id bacteria
  • 5.
  • 6. CLASSIFICATION • International Committee on Taxonomy of Viruses (ICTV) according to morphology and nucleic acid. • Nineteen families • 2 families have RNA genomes • 5 families – envelope • 8 families with DNA genomes - circular genomes • 9 (DNA) - linear genomes
  • 7. • Myoviridae • Contractile tail • Linear ds DNA • T4 phage (T even phage) – E. coli
  • 8. MORPHOLOGY • Tadpole • Hexagonal Head • Cylindrical tail • Head – – Tightly packed core of ds DNA – Surrounded by Protein coat / Capsid • Tail – – hollow core – Contractile sheath surr – Terminal base plate – attached prongs / fibres • Spherical / filamentous • ss DNA / RNA
  • 9.
  • 10.
  • 11.
  • 12.
  • 13. LIFE CYCLE LYTIC PHASE • Virulent • Intracellular multiplication • Bacterial host – lysis • Release of progeny virions LYSOGENIC PHASE • Temperate • Phage DNA combines with host s DNA • Replicating together • No harm to host
  • 14.
  • 15. LYTIC CYCLE • Adsorption • Penetration • Synthesis of phage components • Assembly • Maturation • Release
  • 16.
  • 17. ADSORPTION • Attaches to bacterial surface to area having complimentary receptors to its base plate • Rapid – minutes • Location of bacterial Rs – cell wall / Flagella / Pili • Protoplasts – Not infected ( Transfection )
  • 18. PENETRATION • Lysozyme on tail makes hole on surface • Hollow core pierces the cell wall • Contractile tail sheath propels nucleic acid in bacteria • Initiates synthesis of daughters • Empty head and tail remain outside – GHOST
  • 19.
  • 20.
  • 21.
  • 22. SYNTHESIS OF PHAGE COMPONENTS • First synthesised – Early proteins • Enzymes – manufacture of complex molecules – phage specific • Late proteins – Head and Tail
  • 23. ASSEMBLY • DNA of daughter phage – Packed into head • Tail is added MATURATION • The previous process and conversion to infective stage
  • 24. RELEASE • Lysis of bacterial cell • Phage enzymes • Bacteria becomes spherical during replication of Phage • Defective cell wall • ECLIPSE PHASE – Entry to first intracellular phage formation
  • 25.
  • 26.
  • 27. LYSOGENIC CYCLE • Symbiotic Relationship • Enters the bacterial cell • Integrated in bacterial chromosome • Prophage • Replicates synchronously with host DNA • Lysogeny • Bacterial host metabolism - UNAFFECTED
  • 28. • Lysogenic / Phage conversion – – Prophage confers new properties to bacteria – ? - synthesis of new proteins – Eg – toxin production in Diptheria by beta phage • During repeated multiplication – lysogenic phage may be excised from host chromosome – initiates – Lytic cycle – SPONTANEOUS INDUCTION OF PROPHAGE
  • 29.
  • 30. SPONTANEOUS INDUCTION OF PROPHAGE • UV Rays • Hydrogen peroxide • Nitrogen Mustard • Lysogenic bacteria – Resistant to reinfection – Superinfection Immunity
  • 31. TRANSDUCTION • Carrier of genes between 2 bacteria • 2 Types – Restricted – genes common to bacteria – Generalised – Any • Plasmid mediated drug resistance - MRSA
  • 32.
  • 33.
  • 34. PHAGE ASSAY Zone of lysis after incubation – Plaques Size, shape, nature – characteristic for different phages
  • 35. PHAGE ASSAY • Titrating the number of viable phages • Purification of phages
  • 36. PHAGE TYPING Specificity of Phage bacterium interaction - Id and Typing of Bacteria Adaption of Host – after serial passage in host becomes specific for that host
  • 39.
  • 40.
  • 41.
  • 42.
  • 43.
  • 44.
  • 45.
  • 46.