Bacterial Conjugation - PowerPoint PPT Presentation

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Bacterial Conjugation

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The Tecan test shows that the key plasmid results in the recovery of fluorescence ... The controls show that conjugation is equivalent to a cotransformation of the ... – PowerPoint PPT presentation

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Title: Bacterial Conjugation


1
Bacterial Conjugation
  • Conjugation will be used as a means to transfer
    genetic information from one bacteria to another
  • Cells with a F plasmid or a R plasmid can
    transfer their DNA to other bacteria
  • On the F and R plasmids, DNA transfer begins at
    a specific sequence, OriT, the Origin of
    Transfer.
  • OriT can be knocked out to severely hinder the
    efficiency of conjugation, but the efficiency can
    be restored by a complementation in trans.

2
OriT Complementation
3
Conjugation Assays
Matings with recipient cell ec100Dpir, which
is TriR, show if the F or R plasmid transfers by
the TriK plate and if an OriT plasmid transfers
by the TriA plate
Mutation and complementation of oriT for both F
and R plasmids works!
4
Sending the Key to the Lock
Mating Results of sending the key plasmid to a
cell holding the locked RFP plasmid,
pJ23006-J23077
Mating Results of sending the key plasmid to a
cell holding a OnRFP plasmid, pSB1A2-J01022
Assay shows the key can be transferred
efficiently to the lock!
5
Tecan Fluorescence Assay
The Tecan test shows that the key plasmid results
in the recovery of fluorescence
The controls show that conjugation is equivalent
to a cotransformation of the key and lock
plasmids into a cell
RFP Flurorescence Graph
Graph will NOT BE USED
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