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Bacterial Recombination!

1!

Model Organism E. Coli!

Bacterial colonies, each derived from a single cell!

Figure 5-3!

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Mixing bacterial means mixing genotypes and rare recombinants are observed.!

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Mixing bacterial
genotypes
produces rare
recombinants!

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Figure 5-5b!

If recombination occurs, then it cannot be due to


congugation!

Fine enough to prevent


cells but not DNA or
viruses!
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Bacteria exchange DNA by several processes!

Figure 5-2!

5!

Bacteria conjugate through pili a cell surface appendage, its


synthesis controlled by genes on a small circular molecule
called the F factor.!

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Transfer is not reciprocal: a donar F+ (Fertility) and a


recipient (F-) !

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F plasmids transfer during conjugation using the


rolling circle mechanism the circular F factor
rolls, unwinds one strand of DNA. !

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Two types of DNA transfer can take


place during conjugation!

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The F factor inserts and integrates into


the bacterial chromosome producing a
HFr strain (high frequency
recombination) - because the whole host
chromosome can now replicate and
transfer!

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F factor integrates into the


donar genome changing the
cell into a Hfr cell. !

F factor unrolls, dragging


the donar genome with it
and moves into the host.!

12!

Partial diploid or merozygote!


13!

14!

Episomes exist in 2 states: (1) autonomous (2) integrated!

2 kinds of
transfers,
2 states!

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A single crossover inserts F at a specific locus, which


then determines the order of gene transfer!
Hfr cells rarely convert the
host to Hfr!

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Transfer initiated at OriT, within the F


factor The origin and then chromosomal
DNA is transferred first, the rest of F
factor last.The F factor can orient in
different directions depending on pairing!

The order remains


the same but, the
direction of transfer
may differ,
depending on the
site of integration,
more accurately the
orientation of the F
factor !

17!

Tracking time of marker entry generates a chromosome


map: !
(A) Cross strain 1 and 2 (mix)!
(1)HFr thr+ leu+ azir tonr lac + gal+strs!
(2) F- thr- leu- azis tons lac- gal- strr!
+ = wild,
s = sensitive, !
- = defective r= resistant!
!
(B) Plate onto media containing: !
(1)streptomycin -kills strs cells,!
(2) lacking threonine and leucine!

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Broad Scale Mapping Tracking time of marker entry


generates a chromosome map!
Early - many!

What is the order of


genes ?!

Late -few!

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1) F+ plasmid converts an F- cell to an F+


2) A par6al Hfr foreign chromosome , a fragment crosses
from an a Hfr cell to a recipient cell.
a) Single crossover no recombina6on, fragment degraded
b) Double crossover part of the fragment recombines, the
cross is not reciprocal and the recipient t is recombinant,
whereas the recombinant fragment is degraded

(3) Whole Hfr plasmid crosses intact.
(a)Not integrated or integrate and cleanly excise possibly a
par6al diploid merozygote.
(b)Integrate and excise with a fragment of the neighboring
recipient DNA may carry part of the bacterial plasmid
then it is called an
F plasmid and merozygote.

Fine Scale Mapping Parts of the transferred donor fragment may


be integrated into the host genome through a double crossover!

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Linear fragments are degraded!

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F Plasmid - a plasmid
carrying bacterial DNA.!
!
Produced by outlooping!
!
Cause stable partial
diploids (merozygotes)
in lineages of E coli!

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Transformation bacteria may take fragments of DNA from


their environment and integrate part of them into their
genome, transforming their native genotype if there is
recombination.!
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Transformation: Mechanism of DNA uptake (linear


fragment) by bacteria!

REQUIRES A DOUBLE
CROSSOVER!

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"
Remember, the closer 2 genes, the less likely they will
recombine"
"
If pieces of chromosome do not recombine into the host
chromosome they will be degraded. Most linear
segments of DNA are degraded"
."
A single crossover opens a circular chromosome, 2 (or
an even number) keeps it closed. ."
"
!
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Linear fragments are degraded!

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OTHER EPISOMES
TRANSFERRED DURING
CONGUGATION!

Multiple resistance R- Plasmids.!


A plasmid with segments from many former bacterial
hosts!

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Generalized Transduction!

Structure and function of phage T4!

Figure 5-22!

27!

Electron micrograph of phage infection !

Figure 5-24!

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Cycle of a phage that lyses the host cells!

Figure 5-25!

29!

A phage cross made by doubly infecting the host cell with!


parental phages!

30!

Plaques from recombinant and parental phage progeny!

Figure 5-28!

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Generalized transduction by random incorporation of bacterial !


DNA into phage heads, during phage assembly!

Phages pick up random


pieces of donor DNA!

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From high cotransduction frequencies, close linkage is inferred,!


Alternatively, low cotransduction, distant linkage is inferred!

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Phages integrate into the host genome!

Specialized transduction moves


(transduces) only small segments
on either side of where the
prophage integrates into the host
genome and then drags flanking
areas with it when it excises
(leaves) in a process similar to F
plasmids that incorporate host
DNA!

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