Part:BBa_J33203:Experience

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Applications of BBa_J33203

Team Debrecen-Hungary has decided to give a hand with testing the BBa_J33203 part. As the South-East of Hungary is highly affected by the arsenic problem (there are wells in which the level of arsenic exceeds the allowed limit), we had the possibility to use real-world samples for testing.

We collected samples from two wells which were known to contain elevated arsenic concentration (Szegfű and Madách street, Békéscsaba, Hungary). We used DH5 alpha E. coli cell line with lacZDM15 mutation as host cells, than we prepared a medium consisting of peptone, yeast extract, NaCl, K2HPO4, NaHCO3, bromthymol blue, and autoclaved water samples of different dilutions. We spiked these media with the transformed cells, and after 24 hours of incubation at 37°C we received that the part works also on real world samples, not only on samples made of water and sodium-arsenate. The colour change of Bromothymol Blue seemed to be directly proportional to the concentration of arsenic. The reference dilutions made from Na-arsenate and MilliQ water shows that the concentration of arsenic in our collected samples is at least 15x higher than the WHO recommended limit.

Reference dilutions from Na-arsenate/after 24 hours of incubation. Concentrations from the left to the right and from the top to the bottom: 150 ppb, 100 ppb, 50 ppb, 10 ppb, 5 ppb, 0 ppb, no cell control.

Sample from Szegfű street/after 24 hours of incubation. From the left to the right and from the top to the bottom: MilliQ water control, undiluted sample, 2x dilution, 5x dilution, 10x dilution, 20x dilution, no cell control.

Sample from Madách street/after 24 hours of incubation. From the left to the right and from the top to the bottom: MilliQ water control, undiluted sample, 2x dilution, 5x dilution, 10x dilution, 20x dilution, no cell control.

These samples were used for the publication of the Arsenic Biosensor by the members of the Edinburgh Team.

Reference:

Anal Bioanal Chem. 2011 May;400(4):1031-9. Epub 2011 Mar 27.

A pH-based biosensor for detection of arsenic in drinking water.

de Mora K, Joshi N, Balint BL, Ward FB, Elfick A, French CE., School of Engineering, University of Edinburgh, UK.

For more info about source of the samples or experiments, please contact Lilla Ozgyin (iGEM Team Debrecen_Hungary 2011)

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