Part:BBa_K526002:Design

Design

The last aminoacid (that would form the second aminoacid of the down-stream protein) in the ribo-switch codes for a Threonine (ACU). However, the second aminoacid of GFP is a Serine (AGU). Single base change from C-->G did not significantly alter the functionality of the ribo-switch. Howeve, we could not gurantee the same effect for a huge difference in the second aminoacid.

We were able to see a significant difference in gene expression between 37 degree C and 30 degree C. However, at temperatures less than 30 degree C, there was a severe growth arrest and GFP expression could not be seen both with and with out the riboswitch. This might be due to the lack of arabinose promoter activity at low temperatures like 16 degree.

Reference

Oleksiuk, O., Jakovljevic, V., Vladimirov, N., Carvalho, R., Paster, E., Ryu, William S., Meir, Y., Wingreen, Ned S., Kollmann, M. and Sourjik, V. Thermal Robustness of Signaling in Bacterial Chemotaxis. Cell 145 (2011), pp. 312-321.