Difference between revisions of "Part:BBa K3725120"
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====CHARACTERIZATION==== | ====CHARACTERIZATION==== | ||
Utilizing the phosphate range of 0-150μM and characterization protocol (See: [https://2021.igem.org/Team:Lambert_GA/Pho#:~:text=Engineering%20Success%20page.-,Characterization%20Protocol,-The%20failed%20characterization Modified Protocol]), we characterized the BBa_K3725120 phosphate sensor (Figure 1). However, after overnight incubation, we noticed that the cells produced no fluorescence. Despite resuspending the cells in MOPS media, which contains relatively less phosphate than LB, the part did not exhibit greater fluorescence. Since we confirmed the cloning of the part with successful sequencing results, we concluded that the part was not efficient as a phosphate sensor and discontinued characterization. | Utilizing the phosphate range of 0-150μM and characterization protocol (See: [https://2021.igem.org/Team:Lambert_GA/Pho#:~:text=Engineering%20Success%20page.-,Characterization%20Protocol,-The%20failed%20characterization Modified Protocol]), we characterized the BBa_K3725120 phosphate sensor (Figure 1). However, after overnight incubation, we noticed that the cells produced no fluorescence. Despite resuspending the cells in MOPS media, which contains relatively less phosphate than LB, the part did not exhibit greater fluorescence. Since we confirmed the cloning of the part with successful sequencing results, we concluded that the part was not efficient as a phosphate sensor and discontinued characterization. | ||
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[[File:Fjasksdanfnkcnfkncjak.png|thumb|center|800px|Figure 1. Characterization curve for BBa_K3725120 for phosphate concentrations between 0μM and 150μM. ]] | [[File:Fjasksdanfnkcnfkncjak.png|thumb|center|800px|Figure 1. Characterization curve for BBa_K3725120 for phosphate concentrations between 0μM and 150μM. ]] | ||
Revision as of 20:34, 17 December 2021
PhoB Mutated Consensus Promoter with GFP Reporter
DESIGN
The promoter BBa_K3725130 was designed by replacing parts of BBa_K116401 with the PhoB consensus sequence. A consensus sequence is a calculated order of the most repeated nucleotides found at each position in an alignment of multiple other sequences that have a similar function. By replacing the downstream Pho Regulon with GFP, the part was expected to express fluorescence in the presence of low levels of extracellular phosphate.
CHARACTERIZATION
Utilizing the phosphate range of 0-150μM and characterization protocol (See: Modified Protocol), we characterized the BBa_K3725120 phosphate sensor (Figure 1). However, after overnight incubation, we noticed that the cells produced no fluorescence. Despite resuspending the cells in MOPS media, which contains relatively less phosphate than LB, the part did not exhibit greater fluorescence. Since we confirmed the cloning of the part with successful sequencing results, we concluded that the part was not efficient as a phosphate sensor and discontinued characterization.
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1148