Difference between revisions of "Part:BBa K4218003"
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===Usage and Biology=== | ===Usage and Biology=== | ||
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| + | ==Characterization== | ||
| + | <h2 style="text-align:center; font-family:'Times New Roman', Times, serif;">BBa_K4218003</h2> | ||
| + | <h4 style="font-size:35px; padding-top:20px; font-family:'Times New Roman', Times, serif;">Result</h4> | ||
| + | <h4 style="font-size:35px; padding-top:20px; font-family:'Times New Roman', Times, serif;">The introduction of our experiment:</h4> | ||
| + | <p class="blog-p " style="text-indent:30px; font-size: 20px; font-family:'Times New Roman', Times, serif; text-align:justify; padding-top:20px">We have established a toehold switch that combines with a fluorescent protein, mCherry, on its tail to detect methylated TFPI2 for early screening of non-invasive diagnosis of CRC (https://2021.igem.org/Team:YiYe-China). It is reported that SDC2 methylation and TFPI2 methylation are highly sensitive to CRC (DOI:10.2147/cmar.s375358) . In order to further improve the detection accuracy of CRC compared to last year, we combined the detection of methylated SDC2 with methylated TFPI2 this year. We constructed a toehold switch consisting of the plasmids of double-toehold pSB1C3 (more details were listed below) and double-trigger pCOLADuetTM-1, and then we have a functional verification for the plasmids in the BL21 cells. Finally, we detected methylation SDC2 gene and methylated TFPI2 through the expression of fluorescent protein in cell-free protein expression system.</p> | ||
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Revision as of 11:17, 11 October 2022
T7-double Toehold switch-mcherry
Toehold switch is a special RNA hairpin structure, which contains trigger strand, ribosome binding site, translation start codon and a report gene. Our double toehold switch sequence (BBa_BBa_K4218016) is based on an article called Toehold Switches: De-Novo-Designed Regulators of Gene Expression (Green A, et al., 2014) and the mCherry is added to the downstream of the RNA hairpin structure.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]