Difference between revisions of "Part:BBa K2084002"

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pT7 Toehold

Regulatory ribosome binding site with T7 RNAP promoter.

T7 RNAP-mediated regulatory ribosome binding site that is only translated in the presence of an activating complementary oligonucleotide. The RBS is contained within a hairpin loop that is straightened by the appropriate trigger. As the data below suggest, this toehold switch provides a powerful regulatory element in any circuit. It can be activated by both RNA and DNA, and the expression of the reporter is dependent on the concentration of the toehold switch in a cell-free reaction.

The switch is activated with a complementary RNA strand in the Collins paper. As shown in the figure below, it can also be activated with a complementary DNA strand, though protein expression is lower. Activation is measured by the expression of the reporter lacZ in absorbance at 570 nm.

The activation of the switch also increases with increasing concentration of trigger as shown below. Maximum activation is reached at a concentration of about 100 nM of DNA oligonucleotide trigger.

In PURExpress, an in vitro protein synthesis reaction, maximum expression of the LacZ reporter protein regulated by the switch is reached within 2 hours.

References

Pardee K, Green AA, Ferrante T, Cameron DE, DaleyKeyser A, Yin P, Collins JJ. Paper-based synthetic gene networks. Cell 159 (4): 940-954, 2014.

Sequence and Features