Difference between revisions of "Part:BBa R0080"
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'''Summary:''' We tested expression of this part in ''E.coli'', and corrected the founction description of this part. | '''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: | 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 | https://static.igem.org/mediawiki/2014/f/f9/NUDT_CHINA_2014_K1532008.png | ||
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| − | We draw the curve using the intensity figure of different amount of Arabinose subtracted by the figures of those without Arabinose. | + | 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: | The result is as follows: | ||
Revision as of 03:50, 18 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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 103
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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:
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:
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:
The results indicates that with the existence of Arabinose and AraC, the part BBa_R0080 can activate the down-stream expression observably.