Part:BBa_K1334030:Experience

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••••• Xinyuan Qiu NUDT_CHINA2014

Contribution: iGEM14_NUDT_CHINA Group: iGEM14_NUDT_CHINA/ Author: Xinyuan Qiu Summary: We tested this part in E.coli MATERIAL AND Methods We used the circuit of BBa_K09100 to test the function of this part, the sequence and features of the part BBa_K09100 is as follows: To make the AHL, we transformed the plasmids carrying BBa_K1334030 into the competent cell BL21(DE3). Cultured in liquid LB Media containing 50μg/mL ampicillin in 37 ℃ 220rpm overnight. Then centrifuge the mixture in 13600g for 15min. The supernatant(containing AHL) was used in the fluorometric measurement. We used the supernatant obtained from the culture of clean BL21(DE3)(with no plasmids transformed into) as control. 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 50μg/mL ampicillin to OD0.6, 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. Total time was 710min. The layout of the plate is as follows: Results The result is as follows: As is indicated in the curve above, this part can produce AHL and trigger the Plux as expected.

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