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Bacteriophages
               Dr.T.V.Rao MD




11/20/12       Dr.T.V.Rao MD's Undergraduate Series   1
What are Bacteriophages

     Viruses that attack bacteria were
    observed by Twort and d'Herelle in
   1915 and 1917. T...
11/20/12   Dr.T.V.Rao MD's Undergraduate Series   3
Bacteriophages under Electron
             Microscope




11/20/12     Dr.T.V.Rao MD's Undergraduate Series   4
Bacteriophage (Phage)
 
     Definition - Obligate intracellular parasites
     that multiply inside bacteria by making u...
Bacteriophages:
           Definition & History
                                 
                                     Ba...
BACTRIOPHAGES

    Like most viruses, bacteriophages
      typically carry only the genetic
   information needed for repl...
Bacteriophages:
            Classification
                                   At present, over
                          ...
Bacteriophage
                                    Bacteriophages
                                     make up a
         ...
Bacteriophage




11/20/12     Dr.T.V.Rao MD's Undergraduate Series   10
Bacteriophage
                                    Also known simply as a
                                     phage; a vi...
Composition and
                       Structure
 
         Composition
         − Nucleic            acid               ...
11/20/12   Dr.T.V.Rao MD's Undergraduate Series   13
Phage entering a bacterial cell




11/20/12   Dr.T.V.Rao MD's Undergraduate Series   14
11/20/12   Dr.T.V.Rao MD's Undergraduate Series   15
Bacteriophage showing Lytic and
        lysogenic cycle




11/20/12   Dr.T.V.Rao MD's Undergraduate Series   16
Bacteriophages:
  Virulence Factors Carried On Phage
           
               Temperate phage can go through one of two...
Bacteriophages:
               Lysogenic Conversion
     
           Some lysogenic phage carry genes that
           can...
Bacteriophages
           •
               Used for cloning foreign
               genes among other
               applic...
Bacteriophages
           •
               Used for cloning foreign
               genes among other
               applic...
Bacteriophages
           •
               Once these Phages are isolated and
               recovered they can be used to...
Lytic and Lysogenic cycle




11/20/12         Dr.T.V.Rao MD's Undergraduate Series   22
11/20/12   Dr.T.V.Rao MD's Undergraduate Series   23
11/20/12   Dr.T.V.Rao MD's Undergraduate Series   24
Bacteriophages:
                   Lysogenic Conversion
     Examples of Virulence Factors Carried by Phage
              ...
Lysogenic conversion
In some interactions between lysogenic
phages and bacteria, lysogenic conversion
may occur. It is whe...
Assay for Lytic Phage
                                                             Phage

•
     
       Plaque assay
   ...
Lytic
   vs
Lysogenic
  Cycle




  11/20/12   Dr.T.V.Rao MD's Undergraduate Series   28
Transduction




11/20/12   Dr.T.V.Rao MD's Undergraduate Series   29
Transduction




11/20/12       Dr.T.V.Rao MD's Undergraduate Series   30
Bacterial       Bacterial    Capsid    DNA
       cell wall       chromosome
                                             ...
Tail
                                                                             DNA




4      Maturation:
    Viral com...
Examples:
          * Corynebacterium diphtheria produces the
   toxin of diphtheria only when it is infected by the
 phag...
Medical Applications of Phages
    “I strongly believe phage could become
    an effective antibacterial tool” - Carl Merr...

       The Programme Created by Dr.T.V.Rao MD
       for Undergraduate Learning in Developing
                      Coun...
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Bacteriophages

  1. 1. Bacteriophages Dr.T.V.Rao MD 11/20/12 Dr.T.V.Rao MD's Undergraduate Series 1
  2. 2. What are Bacteriophages Viruses that attack bacteria were observed by Twort and d'Herelle in 1915 and 1917. They observed that broth cultures of certain intestinal bacteria could be dissolved by addition of a bacteria-free filtrate obtained from sewage 11/20/12 Dr.T.V.Rao MD's Undergraduate Series 2
  3. 3. 11/20/12 Dr.T.V.Rao MD's Undergraduate Series 3
  4. 4. Bacteriophages under Electron Microscope 11/20/12 Dr.T.V.Rao MD's Undergraduate Series 4
  5. 5. Bacteriophage (Phage)  Definition - Obligate intracellular parasites that multiply inside bacteria by making use of some or all of the host biosynthetic machinery  Significance − Models for animal cell viruses − Gene transfer in bacteria − Medical applications  Identification of bacteria - phage typing  Treatment and prophylaxsis??? 11/20/12 Dr.T.V.Rao MD's Undergraduate Series 5
  6. 6. Bacteriophages: Definition & History  Bacteriophages are viruses that can infect and destroy bacteria.  They have been referred to as bacterial parasites, with each phage type depending on a single strain of bacteria to act as host. 11/20/12 Dr.T.V.Rao MD's Undergraduate Series 6
  7. 7. BACTRIOPHAGES Like most viruses, bacteriophages typically carry only the genetic information needed for replication of their nucleic acid and synthesis of their protein coats.. They require precursors, energy generation and ribosomes supplied by their bacterial host cell. 11/20/12 Dr.T.V.Rao MD's Undergraduate Series 7
  8. 8. Bacteriophages: Classification  At present, over 5000 bacteriophages have been studied by electron microscopy and can be divided into 13 virus families. 11/20/12 Dr.T.V.Rao MD's Undergraduate Series 8
  9. 9. Bacteriophage  Bacteriophages make up a diverse group of viruses, some of which have complex structures, including double- stranded DNA. 11/20/12 Dr.T.V.Rao MD's Undergraduate Series 9
  10. 10. Bacteriophage 11/20/12 Dr.T.V.Rao MD's Undergraduate Series 10
  11. 11. Bacteriophage  Also known simply as a phage; a virus that attacks and infects bacteria. The infection may or may not lead to the death of the bacterium, depending on the phage and sometimes on conditions. Each bacteriophage is specific to one form of bacteria. 11/20/12 Dr.T.V.Rao MD's Undergraduate Series 11
  12. 12. Composition and Structure  Composition − Nucleic acid Head/Capsid  Genome size  Modified bases Contractile Tail Sheath • Protein −Structure (T ) 4 Size Tail Fibers –  Protection – Head or Infection Base Plate capsid 11/20/12 Dr.T.V.Rao MD's Undergraduate Series 12 – Tail
  13. 13. 11/20/12 Dr.T.V.Rao MD's Undergraduate Series 13
  14. 14. Phage entering a bacterial cell 11/20/12 Dr.T.V.Rao MD's Undergraduate Series 14
  15. 15. 11/20/12 Dr.T.V.Rao MD's Undergraduate Series 15
  16. 16. Bacteriophage showing Lytic and lysogenic cycle 11/20/12 Dr.T.V.Rao MD's Undergraduate Series 16
  17. 17. Bacteriophages: Virulence Factors Carried On Phage  Temperate phage can go through one of two life cycles upon entering a host cell. 1) Lytic: Is when growth results in lysis of the host and release of progeny phage. 2) Lysogenic: Is when growth results in integration of the phage DNA into the host chromosome or stable replication as a plasmid. Most of the gene products of the lysogenic phage remains dormant until it is induced to enter the lytic cycle. 11/20/12 Dr.T.V.Rao MD's Undergraduate Series 17
  18. 18. Bacteriophages: Lysogenic Conversion  Some lysogenic phage carry genes that can enhance the virulence of the bacterial host.  For example, some phage carry genes that encode toxins.  These genes, once integrated into the bacterial chromosome, can cause the once harmless bacteria to release potent toxins that can cause disease. 11/20/12 Dr.T.V.Rao MD's Undergraduate Series 18
  19. 19. Bacteriophages • Used for cloning foreign genes among other applications • Proteins and peptides are fused to the Capsid(surface) of the phage • The combination of the phage and peptide is known as a Fusion Protein 11/20/12 Dr.T.V.Rao MD's Undergraduate Series 19
  20. 20. Bacteriophages • Used for cloning foreign genes among other applications • Proteins and peptides are fused to the Capsid(surface) of the phage • The combination of the phage and peptide is known as a Fusion Protein 11/20/12 Dr.T.V.Rao MD's Undergraduate Series 20
  21. 21. Bacteriophages • Once these Phages are isolated and recovered they can be used to infect bacteria which will create a particle similar to a monoclonal antibody 11/20/12 Dr.T.V.Rao MD's Undergraduate Series 21
  22. 22. Lytic and Lysogenic cycle 11/20/12 Dr.T.V.Rao MD's Undergraduate Series 22
  23. 23. 11/20/12 Dr.T.V.Rao MD's Undergraduate Series 23
  24. 24. 11/20/12 Dr.T.V.Rao MD's Undergraduate Series 24
  25. 25. Bacteriophages: Lysogenic Conversion Examples of Virulence Factors Carried by Phage Gene Bacterium Phage Phenotype Product Vibrio cholerae CTX phage cholerae toxin cholera lambda hemorrhagic Escherichia coli shigalike toxin phage diarrhea clostridial botulinum botulism (food Clostridium botulinum phages toxin poisoning) Corynebacterium corynephage diphtheria diphtheria diphtheriae beta toxin Streptococcus erythrogenic T12 scarlet fever pyogenes toxins 11/20/12 Dr.T.V.Rao MD's Undergraduate Series 25
  26. 26. Lysogenic conversion In some interactions between lysogenic phages and bacteria, lysogenic conversion may occur. It is when a temperate phage induces a change in the phenotype of the bacteria infected that is not part of a usual phage cycle. Changes can often involve the external membrane of the cell by making it impervious to other phages or even by increasing the pathogenic capability of the bacteria for a host. 11/20/12 Dr.T.V.Rao MD's Undergraduate Series 26
  27. 27. Assay for Lytic Phage Phage •  Plaque assay Plaque assay  Method Method – Bacteria Plaque forming unit – +  Plaque forming (pfu) Phage unit (pfu) infectious Measures –  particles Measures infectious particles 11/20/12 Dr.T.V.Rao MD's Undergraduate Series 27
  28. 28. Lytic vs Lysogenic Cycle 11/20/12 Dr.T.V.Rao MD's Undergraduate Series 28
  29. 29. Transduction 11/20/12 Dr.T.V.Rao MD's Undergraduate Series 29
  30. 30. Transduction 11/20/12 Dr.T.V.Rao MD's Undergraduate Series 30
  31. 31. Bacterial Bacterial Capsid DNA cell wall chromosome Capsid Sheath Tail fiber 1 Attachment: Tail Base plate Phage Pin attaches to host cell. Cell wall Plasma membrane 2 Penetration: Phage pnetrates host cell and injects its DNA. Sheath contracted Tail core 3 Merozoites released into bloodsteam from liver may infect new red blood cells 11/20/12 Dr.T.V.Rao MD's Undergraduate Series 31 Figure 13.10.1
  32. 32. Tail DNA 4 Maturation: Viral components are assembled Capsid into virions. 5 Release: Host cell lyses and new virions Tail fibers are released. 11/20/12 Dr.T.V.Rao MD's Undergraduate Series 32 Figure 13.10.2
  33. 33. Examples: * Corynebacterium diphtheria produces the toxin of diphtheria only when it is infected by the phage β. In this case, the gene that codes for the toxin is carried by the phage, not the bacteria. * Vibrio cholerae is a non-toxic strain that can become toxic, producing cholera toxin, when it is infected with the phage CTXφ. * Clostridium botulinum causes botulism. * Streptococcus pyogenes causes scarlet fever. * Shiga toxin 11/20/12 * Tetanus Dr.T.V.Rao MD's Undergraduate Series 33
  34. 34. Medical Applications of Phages “I strongly believe phage could become an effective antibacterial tool” - Carl Merril, Chief of the Laboratory of Biochemical Genetics, National Institute of Mental Health, NIH.  “It might be another string on the bow, such that when (conventional antibiotics) fail, here’s something that has a chance of working. But it’s not going to be a panacea” - Joshua Lederberg, Sackler Foundation Scholar at The Rockefeller University 11/20/12 Dr.T.V.Rao MD's Undergraduate Series 34
  35. 35.  The Programme Created by Dr.T.V.Rao MD for Undergraduate Learning in Developing Countries  Email  doctortvrao@gmail.com 11/20/12 Dr.T.V.Rao MD's Undergraduate Series 35
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