Bacteriophages

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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
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Bacteriophages under Electron Microscope

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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???

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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.
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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.
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Bacteriophages: Classification

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At present, over 5000 bacteriophages have been studied by electron microscopy and can be divided into 13 virus families.

Bacteriophage

Bacteriophages make up a diverse group of viruses, some of which have complex structures, including doublestranded DNA.
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Bacteriophage

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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.
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Composition
Nucleic

Composition and Structure

acid
Head/Capsid

Genome size Modified bases

Structure (T4) Protein

Contractile Sheath

Tail

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Tail Fibers Size Protection Head or Infection capsid Dr.T.V.Rao MD's Undergraduate Series Tail

Base Plate
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Phage entering a bacterial cell

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Bacteriophage showing Lytic and lysogenic cycle

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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.

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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.
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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

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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

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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

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Lytic and Lysogenic cycle

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Bacteriophages: Lysogenic Conversion

Examples of Virulence Factors Carried by Phage Gene
Bacterium Phage Product cholerae toxin Vibrio cholerae CTX phage lambda phage clostridial phages corynephage beta T12 Phenotype cholera hemorrhagic diarrhea botulism (food poisoning) diphtheria scarlet fever

Escherichia coli
Clostridium botulinum Corynebacterium diphtheriae

shigalike toxin
botulinum toxin diphtheria toxin

Streptococcus pyogenes
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erythrogenic toxins

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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.
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Assay for Lytic Phage

Phage

Plaque assay Method Method Plaque forming unit Plaque forming (pfu) unit (pfu) infectious Measures particles Measures infectious particles
Plaque assay

Bacteria + Phage

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Lytic vs Lysogenic Cycle

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Transduction

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Transduction

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Bacterial cell wall

Bacterial chromosome

Capsid

DNA Capsid

Sheath
Tail fiber

1 Attachment:
Phage attaches to host cell.

Base plate Pin Cell wall

Tail

Plasma membrane

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

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Figure 13.10.1

Tail DNA

Maturation: Viral components are assembled into virions.

Capsid

Release: Host cell lyses and new virions are released.

Tail fibers

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Figure 13.10.2

Examples:
* Corynebacterium diphtheria produces the toxin of diphtheria only when it is infected by the phage . 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 CTX. * Clostridium botulinum causes botulism. * Streptococcus pyogenes causes scarlet fever. * Shiga toxin * Tetanus 11/19/2012 Dr.T.V.Rao MD's Undergraduate Series 33

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, heres something that has a chance of working. But its not going to be a panacea - Joshua Lederberg,
Sackler Foundation Scholar at The Rockefeller University
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The Programme Created by Dr.T.V.Rao MD for Undergraduate Learning in Developing Countries

Email

doctortvrao@gmail.com

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