Bacteriophage

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???
 Medical Applications of Phage
• “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

Reassessment of Medicinal Phage Spurs Companies to Study Therapeutic Uses

American Society for Microbiology News 64:620-623, 1998
 Medical Applications of Phage
• Exponential Biotherapies (Rockville, MD)
– Vancomycin resistant Enterococcus facium and Streptococcus
pneumoniae
• Phage Therapeutics (Bothell, WA)
– Staphylococcus aureus and Staphylococcus epidermidis
• Intralytix, Inc. (Baltimore, MD)
– Salmonella in meat and poultry
• Biopharm Ltd. (Tblisi, Georgia)
– Infections associated with burns
• University of Idaho
– Escherichia coli O157:H7 in cattle
Reassessment of Medicinal Phage Spurs Companies to Study Therapeutic Uses. American
Society for Microbiology News 64:620-623, 1998.
Phages eyed as agents to protect against harmful E. coli. American Society for
Microbiology News 65:666-667, 1999.
 Bacteriophage

• T4
• Lambda ()
 Composition and Structure
• Composition
– Nucleic acid Head/Capsid
• Genome size
• Modified bases
– Protein
• Protection
Contractile Tail
• Infection Sheath
• Structure (T4)
Tail Fibers
– Size
Base Plate
– Head or capsid
– Tail
 Infection of Host Cells

• Adsorption
– LPS for T4
• Irreversible attachment
• Sheath Contraction
• Nucleic acid injection
 Types of Bacteriophage

• Lytic or virulent phage: Phage that can only

multiply within bacteria and kill the cell by
lysis. (e.g., T4)
 Lytic Phage Multiplication Cycle

• Eclipse

Number of Infectious Particles

Total Extracellular
Phage Phage
– Early genes
– Phage DNA
synthesis
– Late genes
• Intracellular
Lysis
accumulation Eclipse
Intracellular
accumulatio
n phase
• Lysis and Release
Time after Infection
 Assay for Lytic Phage
Phage

• Plaque assay
Bacteria
– Method +
– Plaque forming unit Phage

(pfu)
– Measures infectious
particles
 Types of Bacteriophage

• Lysogenic or temperate phage: Phage that

can either multiply via the lytic cycle or
enter a quiescent state in the bacterial cell.
(e.g., )
– Expression of most phage genes repressed
– Prophage
– Lysogen
 Events Leading to Lysogeny
• Circularization of the phage chromosome
– Cohesive ends

Cohesive Ends

Lygase

Linear Double Stranded Opened Circle Closed Circle

 Events Leading to Lysogeny

• Site-specific
recombination
– Phage coded
enzyme gal bio

• Repression of the
phage genome
gal
– Repressor protein
bio
– Specific
– Immunity to
superinfection gal bio
 Termination of Lysogeny
• Induction
gal bio
– Adverse conditions
• Role of proteases gal bio

– recA protein
– Destruction of gal

repressor bio

• Gene expression
• Excision
• Lytic growth gal bio
 Lytic vs Lysogenic Cycle?

• Role of repressor
• Role of cro gene product
• Role of proteases
 Significance of Lysogeny

• Model for animal virus transformation

• Lysogenic or phage conversion
– Definition: A change in the phenotype of a
bacterial cell as a consequence of lysogeny
• Modification of Salmonella O antigen
• Toxin production by Corynebacterium diphtheriae