Darn the Luck!

This is Erin reporting after a busy week and a half.

That sneaky cell! All of the work I have done up until this point has not yielded ideal results. Meaning I have not been able to pinpoint a single point mutation that causes temperature sensitivity (Ts) in Burkholderia. However, one clone from my Ts control managed to revert (we think) itself into a non-Ts phenotype. Basically all of the prep work I have done up until this point has not yielded any Ts in Burkholderia thailandensis. Luckily for me one clone seemed to revert from a temperature sensitive phenotype to a non-temperature sensitive one. This may be more exciting than re-creating the Ts phenotype. Now the plan is to verify the clone does actually grow at higher temperatures and sequence the plasmid to see what mutation has occurred. We hope to see a reversion in at least one of the mutations I have been working with during this project. Other options still include testing combinations of mutations to the pUCP28T plasmid.

Electroporation results: None of the transformants exhibit temperature sensitivity. There is still a chance that one of the single mutations could be the culprit but I have not yet been able to generate the mutation in the plasmid by itself.

It will be very interesting to see what becomes of this unexpected mutation.

Until then, happy summer!
Erin

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