Difference between revisions of "Part:BBa K3725120"
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The promoter [https://parts.igem.org/Part:BBa_K3725130 BBa_K3725130] was designed by replacing parts of [https://parts.igem.org/Part:BBa_K116401 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. | The promoter [https://parts.igem.org/Part:BBa_K3725130 BBa_K3725130] was designed by replacing parts of [https://parts.igem.org/Part:BBa_K116401 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: [https://2021.igem.org/Team:Lambert_GA/Pho#:~:text=Engineering%20Success%20page.-,Characterization%20Protocol,-The%20failed%20characterization Modified Protocol]), we characterized the | + | 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. ]] | ||
<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here | ||
Latest revision as of 20:34, 17 December 2021
Status: 500
Content-type: text/html
Software error:
Global symbol "$groups" requires explicit package name at /websites/parts.igem.org/cgi/lib/User.pm line 84. Global symbol "$groups" requires explicit package name at /websites/parts.igem.org/cgi/lib/User.pm line 84. Global symbol "$groups" requires explicit package name at /websites/parts.igem.org/cgi/lib/User.pm line 85. Compilation failed in require at /websites/parts.igem.org/cgi/lib/IconBar.pm line 12. BEGIN failed--compilation aborted at /websites/parts.igem.org/cgi/lib/IconBar.pm line 12. Compilation failed in require at /websites/parts.igem.org/cgi/lib/BBWeb.pm line 9. BEGIN failed--compilation aborted at /websites/parts.igem.org/cgi/lib/BBWeb.pm line 9. Compilation failed in require at /websites/parts.igem.org/cgi/lib/Change.pm line 8. BEGIN failed--compilation aborted at /websites/parts.igem.org/cgi/lib/Change.pm line 8. Compilation failed in require at /websites/parts.igem.org/cgi/lib/Part.pm line 12. BEGIN failed--compilation aborted at /websites/parts.igem.org/cgi/lib/Part.pm line 12. Compilation failed in require at /websites/parts.igem.org/cgi/partsdb/putout.cgi line 8. BEGIN failed--compilation aborted at /websites/parts.igem.org/cgi/partsdb/putout.cgi line 8.
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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.
Status: 500 Content-type: text/html
Software error:
Global symbol "$groups" requires explicit package name at /websites/parts.igem.org/cgi/lib/User.pm line 84. Global symbol "$groups" requires explicit package name at /websites/parts.igem.org/cgi/lib/User.pm line 84. Global symbol "$groups" requires explicit package name at /websites/parts.igem.org/cgi/lib/User.pm line 85. Compilation failed in require at /websites/parts.igem.org/cgi/lib/IconBar.pm line 12. BEGIN failed--compilation aborted at /websites/parts.igem.org/cgi/lib/IconBar.pm line 12. Compilation failed in require at /websites/parts.igem.org/cgi/lib/BBWeb.pm line 9. BEGIN failed--compilation aborted at /websites/parts.igem.org/cgi/lib/BBWeb.pm line 9. Compilation failed in require at /websites/parts.igem.org/cgi/lib/Change.pm line 8. BEGIN failed--compilation aborted at /websites/parts.igem.org/cgi/lib/Change.pm line 8. Compilation failed in require at /websites/parts.igem.org/cgi/lib/Part.pm line 12. BEGIN failed--compilation aborted at /websites/parts.igem.org/cgi/lib/Part.pm line 12. Compilation failed in require at /websites/parts.igem.org/cgi/partsdb/putout.cgi line 8. BEGIN failed--compilation aborted at /websites/parts.igem.org/cgi/partsdb/putout.cgi line 8.
For help, please send mail to this site's webmaster, giving this error message and the time and date of the error.