Revision as of 11:54, 21 October 2019

theophylline riboswitch with optimized RBS + eGFP

This is a part that can test the function of the optimized RBS. The theophylline riboswitch is initially designed by Wieland, M. and Hartig J. S. (2008). We optimize this sequence by replacing the Shine-Dalgarno (SD) sequence with our calculated RBS by thermodynamic model. This switch is theophylline responsive which means this switch will undergo self-cleavage at the presence of theophylline. After self-cleavaging, ribosomes can bind with mRNA and translation will initiate. We use eGFP(BBa_E0040) as our reporter. The fluorescence intensity/OD. 600 can indicate the functionality of this switch. We place this part at the downstream of pBAD promoter. With the induce of L-arabinose, BL21(DE3) cells will transcribe mRNAs containing theophylline switch and eGFP coding sequence. At the presence of theophylline, eGFP protein will be translated and by detecting the fluorescence intensity/OD. 600, the functionality of this switch can be characterized.

Sequence and Features

Background

H2

H3

H4

H5

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