Team:USP-Brazil/Project

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Detecthol

Click on the images below to understand how it works.

Molecular detection

Overview

In order to detect methanol in alcoholic drinks, we searched for P.pastoris promoters inducible by methanol, and preferably known to have well-established genetic recombination techniques.

Otto preparando conteúdo.

The device

Descrição só do device mesmo. Ximena?

Também acho que deveria sair esse título e deixar só o link para o Application:

See the product…

References

Incluir referências do Projeto-Olimpiada: 5, 23 a 27, 29, e a Lin-cereghino acima.

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 		<h4>Overview</h4>
-		<p>The idea of our team this year was to build a biosensor that would respond to methanol by producing a red-coloured protein. In order to develop this biosensor, it is necessary to evaluate the rates at which the promoter Paox1 is activated via methanol or inhibited via ethanol. In addition, we decided to study a modification on the Mxr1p transcription factor, that [Lin-cereghino et al 2006] should alter its interaction with Paox1 by turning ethanol into an activator of the promoter. If the rate of activation by ethanol stays below the rate of activation by methanol, the latter should be identifiable when the drink is diluted. It would then be possible to create a color guide that would help one differ pure and contaminated beverages.</p>
+<p>In order to detect methanol in alcoholic drinks, we searched for P.pastoris promoters inducible by methanol, and preferably known to have well-established genetic recombination techniques.</p>
-		<p>As an alternative, we decided to study another promoter present in <i>Pichia pastoris</i>, namely PFLD1. This second promoter has a high transcription rate when activated, just like Paox1, but instead being activated by methylamine and methanol. Also, it is not repressed by hexoses, like the first one. Since an extensive search in scientific literature did not uncover any data on the regulation of PFLD1 by ethanol, this promoter represents a strong alternative to Paox1, and not only would its characterization be useful for our project, but also it could be very interesting for the expression of heterologous proteins.</p>
+<p style="color:red;><b> >>> tabela só com eles três </b></p>
-<b><h2 style="color:red;">Isso tudo do overview foi feito? Se não, melhor não falar, né?</h2></b>
+<p> The three possible choices (see table above) were PAOX1, PFLD1 and PDAS. Since we did not find much information on PDAS, while PAOX1 and PFLD1 are well characterized, both of them having commercial plasmids for genomic integration [13][14], we chose the latter two as our molecular detectors. That, in a simple genetic construction, could work as a proof of concept. A priori, our output system would be monomeric RFP (mRFP1, Biobrick BBa_E1010), being used for fluorescence tests and for efficiently characterizing the relative promoter strength. We also expect to be able to see the production of RFP with the naked eye, once in E.coli it is shown to be possible.
+.</p>
 		<h4>P<sub>AOX1</sub></h4>