mohammad tabish ahmed - eTheses Repository - University of ...

Chapter 6
can produce products with the wrong orientation, this was checked with restriction digests
and PCR screening. The plasmid was then also sequenced to ensure no undesired mutations
were present before being transformed into strain B to be sent to Pittsburgh. A schematic
representation of the process can be seen in figure 6.1.
6.1.2 Results obtained from America
Since the original complementation assay results could not be replicated in our lab in
Birmingham, the construct had to be sent to Pittsburgh for the complementation analysis.
However, to check the reliability of the experiments the constructs were first sent blind, such
that the collaborators in Pittsburgh were not aware of the identity of the strains. This was
done to see if they could correctly identify the constructs based on their phenotypic results
alone.
Based on the results that were obtained, they were able to correctly identify all the control
strains. Their results showed that both strain A (original strain) and C (complementing
Mscpn60.1) produced mature biofilms (fig 6.2a and 6.2c), while strain B (ΔMscpn60.1)
lacked any signs of biofilm maturation (fig 6.2b).
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