Difference between revisions of "Part:BBa K4656004"

(→‎Usage and Biology)
(→‎Experiment results)
Line 12: Line 12:
 
===Experiment results===
 
===Experiment results===
 
We designed two gene fragments as mentinoned: one directly attaching EGFP downstream of the pchA promoter, and the other appending EGFP after the existing gene segment in enterohemorrhagic E. coli. We will compare the responsiveness and feedback levels of these two fragments to butyrate.
 
We designed two gene fragments as mentinoned: one directly attaching EGFP downstream of the pchA promoter, and the other appending EGFP after the existing gene segment in enterohemorrhagic E. coli. We will compare the responsiveness and feedback levels of these two fragments to butyrate.
−
<html><BR><BR><center><img style="display: block;-webkit-user-select: none;margin: auto;background-color: hsl(0,0%,90%);transition: background-color 300ms;" src="https://static.igem.wiki/teams/4656/wiki/part-plee.jpg"width="466" height="223"></center></html>
+
<html><BR><BR><center><img style="display: block;-webkit-user-select: none;margin: auto;background-color: hsl(0,0%,90%);transition: background-color 300ms;" src="https://static.igem.wiki/teams/4656/wiki/part-plee1.jpg"width="466" height="223"></center></html>
 +
<html><BR><BR><center><img style="display: block;-webkit-user-select: none;margin: auto;background-color: hsl(0,0%,90%);transition: background-color 300ms;" src="https://static.igem.wiki/teams/4656/wiki/part-plee2.jpg"width="466" height="223"></center></html>
  
 
===Reference===
 
===Reference===

Revision as of 13:00, 22 September 2023

PLEE1

PLEE1 is the promoter of the locus of enterocyte effacement which binds to pchA.

Since the specific binding site of pchA and PLEE1 is still unknown, at present, researchers have purchased Escherichia coli genome from atcc and amplified PLEE1 with different fragments by PCR as candidates, and constructed 6 different kinds of plasmids. +1 is the transcription start site TSS.

"Additionally, the best performing plasmid, pC4S-P4, contains a small part of the ler gene in the PLEE1 promoter region."[1] {The pC4S-P4: pDMB,PpchA-pchA-PLEE1(-98+218bp)-gfp;CmR}

Usage and Biology

The Plee promotor is under the control of PchA, we designed and compared Ppcha-pcha-Plee-egfp and Ppcha-pcha-egfp circuit.

Experiment results

We designed two gene fragments as mentinoned: one directly attaching EGFP downstream of the pchA promoter, and the other appending EGFP after the existing gene segment in enterohemorrhagic E. coli. We will compare the responsiveness and feedback levels of these two fragments to butyrate.



Reference

[1]. Schwan W. R. (2011). Regulation of fim genes in uropathogenic Escherichia coli. World journal of clinical infectious diseases, 1(1), 17–25. https://doi.org/10.5495/wjcid.v1.i1.17.

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]