Difference between revisions of "Part:BBa K4218002"

 
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In our project, the trigger sequence is linked to DNA fragment of biomarker by DNA amplification and then was constructed into a plasimd.  After con-transformation into E.coli with the plasmid contain double toehold sequence, the biomarker RNA will also be connected with trigger.  The product with trigger can open the stem-loop structure and form a red fluorescent protein, which is the visible observation result.
 
In our project, the trigger sequence is linked to DNA fragment of biomarker by DNA amplification and then was constructed into a plasimd.  After con-transformation into E.coli with the plasmid contain double toehold sequence, the biomarker RNA will also be connected with trigger.  The product with trigger can open the stem-loop structure and form a red fluorescent protein, which is the visible observation result.
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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.
  
  

Revision as of 10:45, 11 October 2022


This part is composite of T7 promoter, double toehold switch part BBa_K3822000,FP mcherry and termin

In our project, the trigger sequence is linked to DNA fragment of biomarker by DNA amplification and then was constructed into a plasimd. After con-transformation into E.coli with the plasmid contain double toehold sequence, the biomarker RNA will also be connected with trigger. The product with trigger can open the stem-loop structure and form a red fluorescent protein, which is the visible observation result. 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.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]