Difference between revisions of "Part:BBa K1412033"
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Revision as of 09:32, 11 October 2014
Biobrick calable of forming pattern having the mathematical meaning of parabola and hyperbola
What it is
Biobrick K1412002 is the base of this part. We add promoter pTETR in the front of BBa_K20600. In order to realize our goal, we also ligate pCons and TetR to control the express of BBa_K20600 with pTETR.
What it does
When TETR expresses, promoter pTETR can be inhibit. So the lacI can’t work on the promoter pLacI. Our goal gene CheZ can be express, which can recovery chemotaxis of bacteria. But if we add anhydrotetracycline (aTc) in the culture medium, which can restrain the express of TETR, situation can be different. lacI restrain the express of promoter pLacI, the CheZ can’t express.
How to use it
To achieve our goal to form hyperbolic curve and parabola, we use anhydrotetracycline (aTc) and Isopropyl β-D-1-Thiogalactopyranosid (IPTG) as the inhibitor and inductor to regulate to whole circuit.
We use pTET as the substitution for pBAD because when cultivating the bacteria in a long time (about 24 hours) with the inhibitor L-Arabinose, the bacteria may use the L-Arabinose as carbon source. On the contrary, the bacteria will tend to the L-Arabinose. Due to the deficiency caused by the inductor L-Arabinose, we choose aTc to abtain our goal which lead the bacteria tend to oppose direction of the location where is the aTc on the medium plate.
We use M63 semi-solid plate to observe the behavior of E.coli(CL-1). We dot proper concentration of aTc and IPTG with a distance of 1.5 cm. After the two spots dries up, we dot 3μL bacteria on the IPTG, then place the plate at 37℃. By tracking and measuring the chemotaxis diameter of the bacteria on the plate, we get a series of experimental data, more detail in Experimental Data.
Experimental Data
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]