Part:BBa_K3453060
Rosewood DmTrnL-UAA Trigger (Full)
This part is a trigger for sequence-based detection of rosewood. It is a fragment of the TrnL-UAA gene of Dalbergia maritima var. pubescens (BOLD Acc. n° MADA155-14, MADA063-14, MADA062-14, MADA158-14, MADA065-14, MADA104-14, MADA023-14, MADA071-14, MADA110-14) [1].
Usage and Biology
A toehold switch is an RNA–based device containing a ribosome binding site (RBS) and an ATG start codon embedded in a hairpin structure that blocks translation initiation [2]. The hairpin can be unfolded upon binding of a trigger RNA therefore exposing the RBS and the ATG start codon permitting translation of the reporter protein (Figure 1).
Figure 1. Toehold switches principle.
This part is a long trigger for the 6 toehold switches we designed for sequence-based detection of rosewood (BBa_K3453051, BBa_K3453052, BBa_K3453053, BBa_K3453054, BBa_K3453055, BBa_K3453056).
The functionality of this part was tested under controlled of the T7 promoter (BBa_K2150031) and the strong SBa_000587 synthetic terminator (BBa_K3453000) in the composite part BBa_K3453160.
This part proved to be functional: the transcribed rosewood RNA was able to bind to the corresponding toehold switches and a readable output was generated.
Full results are available on the BBa_K3453151, BBa_K3453152, BBa_K3453153, BBa_K3453154, BBa_K3453155, BBa_K3453156 pages in the registry.
References
[1] Hassold S, Lowry PP 2nd, Bauert MR, Razafintsalama A, Ramamonjisoa L, Widmer A. DNA barcoding of Malagasy rosewoods: towards a molecular identification of CITES-listed Dalbergia species. PLoS One (2016) 11, e0157881.
[2] Green AA, Silver PA, Collins JJ, Yin P. Toehold switches: de-novo-designed regulators of gene expression. Cell (2014) 159, 925-939.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 361
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
| None |