Difference between revisions of "Part:BBa K3777025"

Revision as of 18:46, 21 October 2021

KB2-CtcS-T7(ctcO)-3WJdB-PctcS-sgRNA

Basic biosensor device for Chlortetracycline detection.

Usage and Biology
Compared to our another part ctcS-tetM-T7(ctcO)-3WJdB(BBa_K3777018),it adds a binding site which can be regonized by dcas9 proteins and sgRNA.(Fig 1)
When CTR was absent, ctcs would bind to the inducible promoter（PI）and prevent RNA polymerase from initiating transcription, thus repressing the expression of reporter gene. If tet was present, TetR would no longer able to bind to the promoter, resulting in the expression of reporter gene. But BS2 will be regonized and cut by dca9 proteins and sgRNA leading to lower fluorescence expression.
We expressed this circuit in the E. coli BL21(DE3) cells for tetracycline detection. Thus we could roughly deduce the concentration of the antibiotics in the sample according to the fluorescence intensity. Chlortetracycline
Fig.1 Schematic overview of the genetic circuit.

Reference：Alam Khalid K,Tawiah Kwaku D,Lichte Matthew F,Porciani David,Burke Donald H. A Fluorescent Split Aptamer for Visualizing RNA-RNA Assembly In Vivo.[J]. ACS synthetic biology,2017,6(9)
Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 260
Illegal NheI site found at 283
Illegal NheI site found at 1164
Illegal NotI site found at 832
21
INCOMPATIBLE WITH RFC[21]
Illegal XhoI site found at 121
Illegal XhoI site found at 841
Illegal XhoI site found at 1305
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 576

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K3777025 short</partinfo>
-Basic biosensor device for tetracycline detection.
+Basic biosensor device for Chlortetracycline detection.
 <!-- Add more about the biology of this part here-->
 <b><font size="3">Usage and Biology</font></b>
-<br>The genetic circuit was composed of a coding sequence of  tetracycline repressor  which was inserted into an expression vectors with a consitive promoter(BBa_J23114) and RBS(BBa_K3777030), as well as 3WJdB(BBa_K3777000) under the control of T7 promoter (BBa_K3777006).  The terminator we used were BBa_B0010 and BBa_M36305.(Fig 1)
+<br>Compared to our another part ctcS-tetM-T7(ctcO)-3WJdB(BBa_K3777018),it adds a binding site which can be regonized by dcas9 proteins and sgRNA.(Fig 1)
-<br>When tet was absent, TetR would bind to the inducible promoter（PI）and prevent RNA polymerase from initiating transcription, thus repressing the expression of reporter gene. If tet was present, TetR would no longer able to bind to the promoter, resulting in the expression of reporter gene.
+<br>When CTR was absent, ctcs would bind to the inducible promoter（PI）and prevent RNA polymerase from initiating transcription, thus repressing the expression of reporter gene. If tet was present, TetR would no longer able to bind to the promoter, resulting in the expression of reporter gene. But BS2 will be regonized and cut by dca9 proteins and sgRNA leading to lower fluorescence expression.
 <br>We expressed this circuit in the <i>E. coli </i> BL21(DE3) cells for tetracycline detection. Thus we could roughly deduce the concentration of the antibiotics in the sample according to the fluorescence intensity.
 Chlortetracycline