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Recombination at the Molecular Level
• three types
– homologous recombination
– site specific recombination
– transposition
Reciprocal Homologous
Recombination:
Double-Strand Break Model
Nonreciprocal Homologous Recombination: Fox Model

• incorporation of single
strand of DNA into
chromosome, forming
heteroduplex DNA
• thought to occur during
bacterial transformation
Site-specific recombination
• important in insertion of viral genomes into
host chromosomes
• there is only a small region of homology
between inserted genetic material & host
chromosome
transposition & transposable elements
transposable elements = transposons =
mobile genetic elements = “jumping genes”
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replicative
transposition
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R plasmid w/
integrated
Tn3 transposon
What are potential effects of
transposons?
Bacterial Conjugation
• Transfer of genes
between bacteria
that depends on
– direct cell to cell
contact mediated by
a sex pilus
– type IV secretion
system
Evidence for Bacterial Conjugation (Lederberg & Tatum, 1946)
The U-Tube Experiment (Davis, 1950)
Types of Conjugation
1. F+ × F2. Hfr × F3. F’
+

-

F x F Mating
– copy of F factor is transferred to recipient &
does not integrate into the host chromosome
– donor genes usually not transferred
+

-

F xF
Mating
rolling-circle
replication
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Hfr Conjugation
– donor has F factor integrated into its
chromosome (Hfr cell)
– donor genes are transferred to recipient cell
– complete copy of the F factor is usually not
transferred
Figure 14.24a
Figure 14.24b
F’ Conjugation
• F factor incorrectly leaves host chromosome
• some of the F factor is left behind in host
chromosome
• some host genes have been removed along
w/ some of the F factor
– these genes can be transferred to a second host
cell by conjugation
Figure 14.25
Bacterial Transformation
• uptake of naked DNA by a competent cell
followed by incorporation of the DNA into
the recipient cell’s genome
Transduction
• transfer of bacterial genes by viruses
• viruses (bacteriophages) can carry out the
lytic cycle in which the host cell is
destroyed or the viral DNA can integrate
into the host genome, becoming a latent
prophage
Generalized Transduction
• any part of bacterial genome can be
transferred
• occurs during lytic cycle
• during assembly, fragments of host DNA
mistakenly packaged into phage head
– generalized transducing particle
Generalized transduction

abortive transductants = bacteria w/
nonintegrated transduced DNA
Specialized Transduction
• a.k.a. restricted transduction
• carried out only by lysogenic/temperate
phages
• only specific portion of bacterial genome is
transferred
• occurs when prophage incorrectly excises
Specialized/restricted
transduction
Phage lambda
low-frequency
transducing lysate

insert Figure 13.21

high-frequency
transducing lysate
For Tuesday:
Read Ch 17

For Lab Next Week:
Identify unknowns to Group level
Collect materials needed for Team Projects

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Lecture 21

  • 1. Recombination at the Molecular Level • three types – homologous recombination – site specific recombination – transposition
  • 3. Nonreciprocal Homologous Recombination: Fox Model • incorporation of single strand of DNA into chromosome, forming heteroduplex DNA • thought to occur during bacterial transformation
  • 4. Site-specific recombination • important in insertion of viral genomes into host chromosomes • there is only a small region of homology between inserted genetic material & host chromosome
  • 5. transposition & transposable elements transposable elements = transposons = mobile genetic elements = “jumping genes”
  • 6. Please note that due to differing operating systems, some animations will not appear until the presentation is viewed in Presentation Mode (Slide Show view). You may see blank slides in the “Normal” or “Slide Sorter” views. All animations will appear after viewing in Presentation Mode and playing each animation. Most animations will require the latest version of the Flash Player, which is available at http://get.adobe.com/flashplayer.
  • 7.
  • 8.
  • 9. Please note that due to differing operating systems, some animations will not appear until the presentation is viewed in Presentation Mode (Slide Show view). You may see blank slides in the “Normal” or “Slide Sorter” views. All animations will appear after viewing in Presentation Mode and playing each animation. Most animations will require the latest version of the Flash Player, which is available at http://get.adobe.com/flashplayer.
  • 10.
  • 12. Please note that due to differing operating systems, some animations will not appear until the presentation is viewed in Presentation Mode (Slide Show view). You may see blank slides in the “Normal” or “Slide Sorter” views. All animations will appear after viewing in Presentation Mode and playing each animation. Most animations will require the latest version of the Flash Player, which is available at http://get.adobe.com/flashplayer.
  • 14. What are potential effects of transposons?
  • 15.
  • 16. Bacterial Conjugation • Transfer of genes between bacteria that depends on – direct cell to cell contact mediated by a sex pilus – type IV secretion system
  • 17. Evidence for Bacterial Conjugation (Lederberg & Tatum, 1946)
  • 18. The U-Tube Experiment (Davis, 1950)
  • 19. Types of Conjugation 1. F+ × F2. Hfr × F3. F’
  • 20. + - F x F Mating – copy of F factor is transferred to recipient & does not integrate into the host chromosome – donor genes usually not transferred
  • 23. Please note that due to differing operating systems, some animations will not appear until the presentation is viewed in Presentation Mode (Slide Show view). You may see blank slides in the “Normal” or “Slide Sorter” views. All animations will appear after viewing in Presentation Mode and playing each animation. Most animations will require the latest version of the Flash Player, which is available at http://get.adobe.com/flashplayer.
  • 24. Hfr Conjugation – donor has F factor integrated into its chromosome (Hfr cell) – donor genes are transferred to recipient cell – complete copy of the F factor is usually not transferred
  • 25.
  • 28. F’ Conjugation • F factor incorrectly leaves host chromosome • some of the F factor is left behind in host chromosome • some host genes have been removed along w/ some of the F factor – these genes can be transferred to a second host cell by conjugation
  • 30. Bacterial Transformation • uptake of naked DNA by a competent cell followed by incorporation of the DNA into the recipient cell’s genome
  • 31.
  • 32.
  • 33.
  • 34. Transduction • transfer of bacterial genes by viruses • viruses (bacteriophages) can carry out the lytic cycle in which the host cell is destroyed or the viral DNA can integrate into the host genome, becoming a latent prophage
  • 35.
  • 36. Generalized Transduction • any part of bacterial genome can be transferred • occurs during lytic cycle • during assembly, fragments of host DNA mistakenly packaged into phage head – generalized transducing particle
  • 37. Generalized transduction abortive transductants = bacteria w/ nonintegrated transduced DNA
  • 38. Specialized Transduction • a.k.a. restricted transduction • carried out only by lysogenic/temperate phages • only specific portion of bacterial genome is transferred • occurs when prophage incorrectly excises
  • 40.
  • 41. Phage lambda low-frequency transducing lysate insert Figure 13.21 high-frequency transducing lysate
  • 42. For Tuesday: Read Ch 17 For Lab Next Week: Identify unknowns to Group level Collect materials needed for Team Projects