Difference between revisions of "Part:BBa K2996709"
 
(2 intermediate revisions by the same user not shown)
Line 3: Line 3:
 
<partinfo>BBa_K2996709 short</partinfo>
 
<partinfo>BBa_K2996709 short</partinfo>
  
−
LuxI is put downstream of BBa_K2996705, as a reporter, which is a synthetase for acyl-homoserine lactones (AHL).
+
LuxI is put downstream of BBa_K2996704, as a reporter, which is a synthetase for acyl-homoserine lactones (AHL).  
−
dcas9 binding to lure means the occurrence of off-target incidence. RpoA, which is linked to dcas9, will recruits RNA polymerase and the gene downstream of the promotor will be activated.  
+
We develop this to record off-target incidence. When the dcas9-sgRNA complex binds to target sequence, RpoA will recruit RNA polymerase, promoter J23117 will be activated. Then, by measuring the biological activity of the downstream reporter gene, we will know the effectiveness of this transcription activation device under different conditions.
−
We develop a way to record off-target incidence, with LuxI gene under the control of promotor J23117. Increased expression of luxI enzyme will synthesis enough AHL molecules, which can penetrate to the bacteria capable of information storage.  
+
  
 +
Increased expression of luxI enzyme indicates the off-target incidence, and AHL molecules, synthesized by LuxI, will penetrate to the bacteria capable of information storage.
  
 +
<center>{{#tag:html|<img style="max-width: 80%" src="https://2019.igem.org/wiki/images/7/73/T--SJTU-BioX-Shanghai--wet_lab-IF1.1.jpg" alt="" />}}</center>
 +
 +
<center><b>Figure 1.</b> <i>Gene circuit for transcription activation and information storage based on AHL</i> </center>
  
  

Latest revision as of 14:48, 21 October 2019


LuxI downstream of activation unit containing lure

LuxI is put downstream of BBa_K2996704, as a reporter, which is a synthetase for acyl-homoserine lactones (AHL). We develop this to record off-target incidence. When the dcas9-sgRNA complex binds to target sequence, RpoA will recruit RNA polymerase, promoter J23117 will be activated. Then, by measuring the biological activity of the downstream reporter gene, we will know the effectiveness of this transcription activation device under different conditions.

Increased expression of luxI enzyme indicates the off-target incidence, and AHL molecules, synthesized by LuxI, will penetrate to the bacteria capable of information storage.

Figure 1. Gene circuit for transcription activation and information storage based on AHL


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 52
    Illegal NheI site found at 75
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 743
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 39
  • 1000
    COMPATIBLE WITH RFC[1000]