Generator

Part:BBa_K1709003:Experience

Designed by: Astrid Deryckere   Group: iGEM15_KU_Leuven   (2015-09-16)
Revision as of 16:10, 26 September 2015 by AstridD (Talk | contribs)

To characterize the CheZ-GFP BioBrick, the fragment containing a RBS was cloned directly after a strong promotor (BBa_J23101). The first figure shows the gel right before ligation.
The colonies were checked by restriction mapping using BcuI and PstI (results not shown). The DNA sequence was also confirmed by DNA sequencing. Results can be provided by email. The presence of colonies expressing GFP proves that the plasmid was designed and cloned correctly.

Further DNA sequencing proved the correct sequence, as designed.

RBS-CheZ seq.JPG

Part of the sequence alignment with on top the reference sequence and on the bottom the sequencing results.

KU Leuven GelPurification.jpeg

Gel after purification. Lanes 2-5: insert (1400bp). Lane 6: linearized vector. Lanes 7-10 : insert. Lane 11: linearized vector


KU Leuven fluorescence.jpg

GFP is expressed in the cells. This confirms the correct construction of the BioBrick


Further characterization could be done by transforming the cheZ knockout Keio strain with this plasmid. These cells should then regain their possibility to swim.



Applications of BBa_K1709003

Motility control

User Reviews

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