Team:Groningen/Notebook/tasA Transformation t49
Blue Bio Energy
tasA Transformation in Bacillus (t49)
tasA was transformed into Bacillus to improve its biofilm properties.
Our goal is to overexpress TasA by adding a second copy of the tasA gene under the control of a salt inducible promoter. This overexpression will lead to a stronger biofilm when the biofilm is exposed to salt water.
The transformation was successful, but it was found out that instead of tasA RFP was present.
Transformation
09:24, 20 July 2015 - 16:30, 20 July 2015
Sample 33 (tasA with Psalt in K823023) was transformed into Bacillus subtilis 3610 comI. For this a comI colony was taken from a plate and put in a tube with 2 mL MC 1x. The tube was put in the shaking incubator for ∼5 hours. After 5 hours of growing the OD at 600 nm was seen to be 0.15. 400 µL of culture was added to a new tube, and 5 µL of plasmid DNA (sample 33) was added. The new tube was placed in the shaking incubator at 37 °C.
00:00, 22 July 2015 - 00:00, 22 July 2015
The sequencing results showed that the insert into the K823023 backbone was not the correct one. RFP was inserted instead of tasA with the salt promoter. The transformed Bacillus was discarded.