Team:Newcastle/26 August 2010

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Screening Bacillius subtilis transformants

Aim

The aim of the experiment is to identify those colones that have the plasmid integrated at the correct position in the chromosome, which is the amyE locus. Those that have integrated at the correct position will not be able to break down starch, which can be tested by exposing the colonies on the starch plates to iodine.

Results

None of our colonies had halos, therefore the transformation and integration was successful.


Starchplate.jpg
Starchplate2.jpg

However when we checked the colonies under a microscope the cells were filamentous due to integration elsewhere on the Bacillus subtilis 168 chromosome perhaps at the native yneA locus.

Filamentous cells
Filamentous cells showing GFP signal
Normal Bacillus subtilis 168



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