Difference between revisions of "Part:BBa K2120425"

Line 3: Line 3:
 
<partinfo>BBa_K2120425 short</partinfo>
 
<partinfo>BBa_K2120425 short</partinfo>
  
This is one of our application circuit.
+
This circuit consists of two parts:the inhibitor device and reporter device ,which have already been made into basic parts of biobricks.
 +
We choose three different promoters (BBa_J23109,BBa_J23116,BBa_J23106) whose reported strengths are respectively low, medium and strong from constitutive promoter family to express the inhibitor tetR and cI ,
 +
a reporter protein-mRFP (BBa_E1010) was used as a signal to represent if the concentration of inhibitor controlled by different promoters can repress the expression of downstream gene controllled by inducible promoter completely .At first,we test fluorescence intensity of mRFP protein expressed by each circuit in high copy plasmid pSB1K3 ,the results show that inhibitor devices with J23116 and J23109 cannot inhibit the expression of mRFP ,which means that the strength of these two promoters is too weak to turn off the reporter device in our circuit.
  
Our application circuit circuit consists of two parts:the inhibitor device and killer device ,which have already been made into basic parts of biobricks.
 
 
We choose four different promoters whose reported strengths are respectively low, medium,strong and the strongest from constitutive promoter family to express the inhibitor tetR and cI ,and then we construct three plasmid with different copy number containing this circuit, in order to find out the proper promoter which meets the threshold that trigger the killer system.
 
 
Go our [https://parts.igem.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2016&group=BIT-China Parts Registry] to learn more.
 
  
  

Revision as of 08:45, 20 October 2016


B0015+cI+B0034+J23106+pR+B0032+mRFP+B0015

This circuit consists of two parts:the inhibitor device and reporter device ,which have already been made into basic parts of biobricks. We choose three different promoters (BBa_J23109,BBa_J23116,BBa_J23106) whose reported strengths are respectively low, medium and strong from constitutive promoter family to express the inhibitor tetR and cI , a reporter protein-mRFP (BBa_E1010) was used as a signal to represent if the concentration of inhibitor controlled by different promoters can repress the expression of downstream gene controllled by inducible promoter completely .At first,we test fluorescence intensity of mRFP protein expressed by each circuit in high copy plasmid pSB1K3 ,the results show that inhibitor devices with J23116 and J23109 cannot inhibit the expression of mRFP ,which means that the strength of these two promoters is too weak to turn off the reporter device in our circuit.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 941
    Illegal NheI site found at 964
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1614
    Illegal AgeI site found at 1726
  • 1000
    COMPATIBLE WITH RFC[1000]