Difference between revisions of "Part:BBa R0080"

Revision as of 01:54, 19 October 2014

Promoter (AraC regulated)

AraC operator, truncated to include araO1, araI1, araI2, c-amp1, and c-amp2 sites. This operator should *activate* transcription in the presence of AraC or the simple sugar [http://openwetware.org/wiki/Arabinose arabinose]

b/c the operator lacks the araO2 site, there should not be araC-mediated repression.

Usage and Biology

araC binding region from E.coli

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 103
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 18: / Line 18: @@
-==Contribution: iGEM14_NUDT_CHINA==
-'''Group:''' iGEM14_NUDT_CHINA/ '''Author:''' Xinyuan Qiu
-'''Summary:''' We tested expression of this part in ''E.coli'', and corrected the founction description of this part.
-===Materials and Methods===
-We used the circuit of BBa_K1532008 to test the function of this part, the sequence and features of the part BBa_K1532008 is as follows:
-https://static.igem.org/mediawiki/2014/f/f9/NUDT_CHINA_2014_K1532008.png
-Fluorometric measurement were processed with Fluoroskan(R) Ascent FL from ThermoTM using the filters of 485nm and 538nm. The E.coli to be tested was cultured in liquid LB media containing 35μg/mL chloramphenicol to OD1.6, than split into 96 well plates (from ThermoTM) by 150μL/well. The plate was then put into the Fluoroskan(R) Ascent FL and cultured in 25℃ and background shake of 90rpms. The Fluorometric measurement was processed with the interval time of 10 minutes.
-The layout of the plate is as follows:
-https://static.igem.org/mediawiki/2014/5/50/NUDT_CHINA_R0080_CON1.jpg
-We draw the curve using the intensity figure of different amount of Arabinose subtracted by the figures of those without Arabinose.
-===Results===
-The result is as follows:
-https://static.igem.org/mediawiki/2014/8/89/NUDT_CHINA_2014_RCC80_CON2.jpg
-The results indicates that with the existence of Arabinose and AraC, the part BBa_R0080 can activate the down-stream expression observably.
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