Based on reverse engineering principles, we engineered our bacteria into valid bioreporters for Hg(II) in aquatic environment. Rational design of genetic circuit topology was conducted to confer mercury sensor and regulator components high efficiency and robustness. Additionally, a new approach based on the use of multiple bioreporter cell lines to control assay variations and to improve in field application ease was also developed.
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Based on reverse engineering principles, we<br>
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engineered our bacteria into valid bioreporters <br> for Hg(II) in aquatic environment. Rational desi-<br> -gn of genetic circuit topology was conducted to <br> confer mercury sensor and regulator compone-<br> -nts high efficiency and robustness. Additionally,<br> a new approach based on the use of multiple <br> bioreporter cell lines to control assay variations <br> and to improve in field application ease was also <br> developed. <br>
MerR family TFs share a high homology at their <br> metal binding domains, which implies that our strategies of bioreporter <br> and bioabsorbent engineering might be applicable to other cases. Then our<br> reverse engineering strategy was expanded to another common toxic heavy<br> metal, lead (Pb) to prove the validness of our engineering strategy.<a href="https://2010.igem.org/Team:Peking/Project/Expansion"><font size=2>learn more</font></a></p>
A closer look into mercury- and lead- responsive regulators was conducted via 3D structure modeling, followed by rational design of metal binding peptides (MBP) for various heavy metals. Mercury and lead MBPs were accomplished in our bioware experiment. Then high-performance whole-cell bioabsorbent was constructed by expression of MBP on outer membrane surface, periplasm and cytosol of E.coli cells, verified by following function test.
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<p> A closer look into mercury- and lead-responsive<br> regulators was conducted via 3D structure mode-<br> -ling, followed by rational design of metal binding <br> peptides (MBP) for various heavy metals. Merc-<br> -ury and lead MBPs were accomplished in our<br> bioware experiment. Then high-performance <br> whole-cell bioabsorbent was constructed by <br> expression of MBP on outer membrane surface, <br> periplasm and cytosol of E.coli cells, verified by<br> following function test.
MerR family TFs share a high homology at <br> their metal binding domains, which implies that our strategies of bioreporter and bioabsorbent engineering might be applicable to other cases. Then our reverse engineering strategy was expanded to another common toxic heavy metal, lead (Pb) to prove the validness.
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<br><p> <font size=4>Application:</font> Traditional bioreporters are not practical when it comes to <br> in field application. One crucial factor that restricts the in field application is the<br> preservation condition of the bacteria, often requiring costly instrument.We <br> endeavored to solve this hard truth by development of a standard preservation<br> method which endows our bioreporters with application ease. <a href="https://2010.igem.org/Team:Peking/Project/Application">learn more</a></p>
Traditional bioreporters are not practical when it <br>comes to in field application. One crucial factor is the preservation condition of the bacteria, often requiring costly instrument. We endeavo-<br>-red to solve this hard truth by development of a standard preservation method which endows our bioreporters with application ease.
Biosensor:
Based on reverse engineering principles, we engineered our bacteria into valid bioreporters for Hg(II) in aquatic environment. Rational design of genetic circuit topology was conducted to confer mercury sensor and regulator components high efficiency and robustness. Additionally, a new approach based on the use of multiple bioreporter cell lines to control assay variations and to improve in field application ease was also developed.
Bioabsorbent:
A closer look into mercury- and lead- responsive regulators was conducted via 3D structure modeling, followed by rational design of metal binding peptides (MBP) for various heavy metals. Mercury and lead MBPs were accomplished in our bioware experiment. Then high-performance whole-cell bioabsorbent was constructed by expression of MBP on outer membrane surface, periplasm and cytosol of E.coli cells, verified by following function test.
Expansion:
MerR family TFs share a high homology at their metal binding domains, which implies that our strategies of bioreporter and bioabsorbent engineering might be applicable to other cases. Then our reverse engineering strategy was expanded to another common toxic heavy metal, lead (Pb) to prove the validness.
Application:
Traditional bioreporters are not practical when it comes to in field application. One crucial factor is the preservation condition of the bacteria, often requiring costly instrument. We endeavo- -red to solve this hard truth by development of a standard preservation method which endows our bioreporters with application ease.
Parts
Our team submitted a library of thoroughly characterized and standardized parts. Therefore contributing an alternative set of tools towards heavy metal detection and decontamination in the iGEM context. What's more, as our MerR-family-derivated engineering method could be applicable in nearly all cases of heavy metal, our parts and corresponding documents provide a streamlined method for heavy metal bioreporter and bioabsorbent construction.
Eighteen students and three instructors are working on our project during this summer. We split up into several subgroups whose focus and results you can follow on the Notebook or Project pages. If you want to know more about the subgroups and the people involved, meet us on our Team page.
It's an unforgettable summer, during which we spared no effort to fight for our dreams and the honour of Peking iGEM team. Photos recorded every footprint of everyone. Join us together!
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