Difference between revisions of "Part:BBa K2550000:Experience"
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| − | <i>Figure 3: Image showing the sequencing results of the T7 Toehold LacZ construct. Annotated in a white arrow is the biobricked T7 promoter (BBa_I719005); this is followed by the biobricked toehold sequence which was used in the NuPack software to illustrate the functioning Toehold Switch (BBa_K2550100). Illustrated in the maroon arrow is the biobricked LacZ gene (BBa_I732005) sequence that | + | <i>Figure 3: Image showing the sequencing results of the T7 Toehold LacZ construct. Annotated in a white arrow is the biobricked T7 promoter (BBa_I719005); this is followed by the biobricked toehold sequence which was used in the NuPack software to illustrate the functioning Toehold Switch (BBa_K2550100). Illustrated in the maroon arrow is the biobricked LacZ gene (BBa_I732005) sequence that justified that the one base pair wobble mutation was successful.</i> |
===User Reviews=== | ===User Reviews=== | ||
Revision as of 19:22, 16 October 2018
Applications of BBa_K2550000
Figure 1: Toehold structure implemented in the T7 Toehold LacZ biobrick that was developed in the Nupack software which confirmed the shape of the toehold sequence.
The toehold sequence, part BBa_K2550100, is 108 bp long and was obtained from the 144 first generation orthogonal toehold switches collection from the 2017 Collins paper titled Toehold Switches: De-Novo-Designed Regulators of Gene Expression. This distinct riboregulator follows the T7 promoter that is assembled in the T7 Toehold LacZ biobrick. This image above is a visual representation of this unique toehold structure.
Figure 2: The T7 Toehold LacZ biobrick transformed on a chloramphenicol and xgal plate expressing blue pigment without trigger sequence as a result of the strong T7 promoter (Image on the left). T7 Toehold LacZ and trigger sequence transformed in a dual plasmid transformation on a carbenicillin, chloramphenicol, and xgal plate that is expressing a blue pigment due to presence of trigger sequence (Image on the right).
T7, part BBa_I719005, is a strong constitutive promoter derived from the T7 bacteriophage that expresses proteins in the presence of T7 polymerase. It was observed that the toehold expressed a blue pigment when inoculated into xgal and Luria Broth. Although a lighter shade than when fully induced, we hypothesize that this pigment is due to toehold leakiness as a result of the strength of the T7 promoter.
Sequencing Analysis
Figure 3: Image showing the sequencing results of the T7 Toehold LacZ construct. Annotated in a white arrow is the biobricked T7 promoter (BBa_I719005); this is followed by the biobricked toehold sequence which was used in the NuPack software to illustrate the functioning Toehold Switch (BBa_K2550100). Illustrated in the maroon arrow is the biobricked LacZ gene (BBa_I732005) sequence that justified that the one base pair wobble mutation was successful.
User Reviews
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