Team:Tokyo-Nokogen/Project

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Latest revision as of 03:57, 22 October 2009

The production of recombinant protein is an important step to many experiments. Unfortunately, procedures can often be quite laborious and annoying (e.g., centrifugation, sonication,…). Our goal is to use synthetic biology to provide an easy solution. We named our system the Escherichia coli auto protein synthesizer (ESCAPES). It takes advantage of four concepts: 1) light switches, 2) cellular self-aggregation, 3) autolysis, and 4) transcriptional signal counter. Click on the labels in the figure below for further details. With our ESCAPES system, we would only need to irradiate the cells twice to produce a crude extract of our target protein.

The first induction with green light induces the expression of antigen 43, which causes the E.coli to self-aggregate and settle to the bottom. The medium is then decanted and replaced with a small volume of buffer. The aggregated cells are once again irradiated with green light, inducing the expression of holin and endolysin, which causes the autolysis of the bacterial cells. ESCAPES will allow us to escape from the annoying centrifugation and lysis steps, to conveniently produce crude extracts of our target protein.

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-<td>&nbsp;</td>
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-<h3><p style="margin-left:50px; margin-right:50px"> In biochemistry experiments, expression and getting objective protein is popular. But the procedure is almost vexatious complication due to need many solutions and centrifugations. So we hope to solve this trouble to construct alternative easily system with synthetic biology. The system we named <I>Escherichia coli</I> auto protein synthesizer. It has three components. 1. light switches 2. Aggregation 3. Signal count system. A figure that shows procedures this system is below. We only have to irradiate <I>E.coli</I> three times and purification to get proteins in it.<h3>
+<h3><p style="margin-left:50px; margin-right:50px"> The production of recombinant protein is an important step to many experiments. Unfortunately, procedures can often be quite laborious and annoying (e.g., centrifugation, sonication,…). Our goal is to use synthetic biology to provide an easy solution. We named our system the <I>Escherichia coli</I> auto protein synthesizer (ESCAPES). It takes advantage of four concepts: 1) light switches, 2) cellular self-aggregation, 3) autolysis, and 4) transcriptional signal counter. Click on the labels in the figure below for further details. With our ESCAPES system, we would only need to irradiate the cells twice to produce a crude extract of our target protein.
 <table width="730" height="600" border="0" style="background:url(https://static.igem.org/mediawiki/2009/4/48/ESCAPeS1.png) no-repeat; margin-left:50px">
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-<h3><p style="margin-left:50px; margin-right:50px"> By first induction with red light, objective protein is expressed in <I>E.coli</I>. Then same induction repeated, but Antigen 43 that is protein for aggregation <I>E.coli</I> is expressed by signal count system. Once medium replaced, we think that we use blue light with phycoerythrine as receptor. Thereby, holin and endolysin are expressed and lysis <I>E.coli</I>. Finally, by purification of objective protein, this system is completed.<h3>
+<p style="margin-left:50px; margin-right:50px"> The first induction with green light induces the expression of antigen 43, which causes the <I>E.coli</I> to self-aggregate and settle to the bottom. The medium is then decanted and replaced with a small volume of buffer. The aggregated cells are once again irradiated with green light, inducing the expression of holin and endolysin, which causes the autolysis of the bacterial cells. ESCAPES will allow us to escape from the annoying centrifugation and lysis steps, to conveniently produce crude extracts of our target protein.
 <p align="center"><a href="https://2009.igem.org/Team:Tokyo-Nokogen/Project">