Difference between revisions of "Part:BBa K2120416"

Revision as of 08:43, 20 October 2016

B0015+tetR+B0034+J23119+pTet+B0034+mRFP+B0015

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.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 851
Illegal NheI site found at 874
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 1528
Illegal AgeI site found at 1640
1000
COMPATIBLE WITH RFC[1000]

@@ Line 5: / Line 5: @@
 This is one of our application 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.
+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.
-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.