Difference between revisions of "Part:BBa K2660010"

Revision as of 02:30, 18 October 2018

glucose-responsive regulatory circuit

A complete glucose-responsive regulatory circuit (pGRS) for gram-positive bacteria. This device was built in Bacillus subtilis and tested with glucose, fructose, and lactose. It activates the gene expression in presence of glucose or fructose and deactivates in lactose.

Experiments We grew B. subtilis carrying plasmid pGRS in LB media supplemented with 2% fructose and lactose overnight. The cultures were centrifuged, the cell pellet was resuspended in NaCl 0,9%, and we measured the absorbance at 600 nm and fluorescence of mCherry. Figure 2 shows results for the positive control (B. subtilis producing mCherry constitutively) and the test strain (B. subtilis carrying the synthetic circuit). B. subtilis 168 was used as negative control and its intrinsic cell fluorescence was considered as blank.

Our Glucose Responsive System works as expected. Gene expression is ON when glucose/fructose is present, and it is completely OFF in the absence of it. This is a very important result to show the specificity of our circuit and to demonstrate that we accomplished our goal of avoiding leakages with this design.

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]

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 <partinfo>BBa_K2660010 short</partinfo>
-A complete glucose-responsive regulatory circuit for gram-positive bacteria. This device was built in <i>Bacillus subtilis</i> and tested with glucose, fructose, and lactose. It activates the gene expression in presence of glucose or fructose and deactivates in lactose.
+A complete glucose-responsive regulatory circuit (pGRS) for gram-positive bacteria. This device was built in <i>Bacillus subtilis</i> and tested with glucose, fructose, and lactose. It activates the gene expression in presence of glucose or fructose and deactivates in lactose.
+<b>Experiments</b>
+We grew B. subtilis carrying plasmid pGRS in LB media supplemented with 2% fructose and lactose overnight. The cultures were centrifuged, the cell pellet was resuspended in NaCl 0,9%, and we measured the absorbance at 600 nm and fluorescence of mCherry. Figure 2 shows results for the positive control (B. subtilis producing mCherry constitutively) and the test strain (B. subtilis carrying the synthetic circuit). B. subtilis 168 was used as negative control and its intrinsic cell fluorescence was considered as blank.
+[[File:T--Unesp Brazil--graficonathan.png]]
+Our Glucose Responsive System works as expected. Gene expression is ON when glucose/fructose is present, and it is completely OFF in the absence of it. This is a very important result to show the specificity of our circuit and to demonstrate that we accomplished our goal of avoiding leakages with this design.
 <!-- Add more about the biology of this part here