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Team:WHU-China/Project/Death
From 2012.igem.org
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<h2> Outline</h2> | <h2> Outline</h2> | ||
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| - | We plan to place endonuclease and YhjH gene downstream the xylR | + | We plan to place endonuclease and YhjH gene downstream the xylR induced promoter to make bacteria die and lose their adhesion when exposed to xylose. |
</p> | </p> | ||
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| - | In order to terminate the overreaction of fatty acid metabolism and cellulose synthesis, we design an elaborate device | + | In order to terminate the overreaction of fatty acid metabolism and cellulose synthesis, we design an elaborate device. Firstly, we determined to use endonuclease BglII(BBa_K112106, a protein |
| - | + | ||
| - | + | ||
which can kill the microbe without inducing cell lysis. Since AdrA but not YhjH contains this restriction site, our E.coslim will lose their adhesion more easily. To control the expression of endonuclease, a regulator is | which can kill the microbe without inducing cell lysis. Since AdrA but not YhjH contains this restriction site, our E.coslim will lose their adhesion more easily. To control the expression of endonuclease, a regulator is | ||
| - | necessary. We | + | necessary. We selected xylose as a controller because it is more healthy and hard to be absorbed by human beings. We found an ideal |
promoter BBa_K625002, which can be activated by the transcriptional regulator XylR. We hope that eating xylose will | promoter BBa_K625002, which can be activated by the transcriptional regulator XylR. We hope that eating xylose will | ||
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In addition, we use a pair of genes to strengthen the accuracy of controlling, namely YhjH. YhjH gene will counteract the | In addition, we use a pair of genes to strengthen the accuracy of controlling, namely YhjH. YhjH gene will counteract the | ||
| - | impact of AdrA. YhjH protein can convert c-di-GMP | + | impact of AdrA. YhjH protein can convert c-di-GMP into GMP. Thus cells will lose their adhesion and become vulnerable to |
endonuclease. | endonuclease. | ||
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is highly necessary. | is highly necessary. | ||
| - | To prevent horizontal gene transfer, we designed a two plasmids system. In one plasmid, xlyR repressor encoding gene from | + | To prevent horizontal gene transfer, we designed a two-plasmids system. In one plasmid, xlyR repressor encoding gene from |
Bacillus subtilis will be cloned. On the other plasmid, metabolic gene will be coupled with xylR repressed death system. In | Bacillus subtilis will be cloned. On the other plasmid, metabolic gene will be coupled with xylR repressed death system. In | ||
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our GMOs, since death system is suppressed by xylR in low xylose concentration, the bacteria will not die. However, when | our GMOs, since death system is suppressed by xylR in low xylose concentration, the bacteria will not die. However, when | ||
| - | the HGT of those high efficient metabolic genes | + | the HGT of those high efficient metabolic genes happens, the death system will not be suppressed, and the recipient will |
be killed. | be killed. | ||
</p> | </p> | ||
</html> | </html> | ||
Revision as of 19:03, 23 September 2012
Death Device
Outline
We plan to place endonuclease and YhjH gene downstream the xylR induced promoter to make bacteria die and lose their adhesion when exposed to xylose.
Description
In order to terminate the overreaction of fatty acid metabolism and cellulose synthesis, we design an elaborate device. Firstly, we determined to use endonuclease BglII(BBa_K112106, a protein
which can kill the microbe without inducing cell lysis. Since AdrA but not YhjH contains this restriction site, our E.coslim will lose their adhesion more easily. To control the expression of endonuclease, a regulator is
necessary. We selected xylose as a controller because it is more healthy and hard to be absorbed by human beings. We found an ideal
promoter BBa_K625002, which can be activated by the transcriptional regulator XylR. We hope that eating xylose will
subsequently induce the expression of endonuclease, ending up with the death of our E.coslim.
In addition, we use a pair of genes to strengthen the accuracy of controlling, namely YhjH. YhjH gene will counteract the
impact of AdrA. YhjH protein can convert c-di-GMP into GMP. Thus cells will lose their adhesion and become vulnerable to
endonuclease.
Forestalling HGT
In human gut, there are billions of microbes. Horizontal gene transfer of those high efficient metabolic genes between GMOs and normal intestine residents may get out of control and cause disastrous effects. Therefore, a mechanism to prevent HGT is highly necessary. To prevent horizontal gene transfer, we designed a two-plasmids system. In one plasmid, xlyR repressor encoding gene from Bacillus subtilis will be cloned. On the other plasmid, metabolic gene will be coupled with xylR repressed death system. In our GMOs, since death system is suppressed by xylR in low xylose concentration, the bacteria will not die. However, when the HGT of those high efficient metabolic genes happens, the death system will not be suppressed, and the recipient will be killed.
