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Team:IvyTech-South Bend/Project
From 2012.igem.org
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== '''Overall project''' == | == '''Overall project''' == | ||
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== Project Details== | == Project Details== | ||
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We compared the sensitivity of J33201 to the Gronigen Team’s K190015 pArs site using the Endy Lab’s expression vector J61002. We loaded J33201 into a kanamycin resistant backbone pSB1K3 and exchanged it with K190015 by perfoming an upstream cut of both to leave the RFP in the J61002 backbone. (Ratio of J33201 to K190015 plasmid DNA was 3:1) | We compared the sensitivity of J33201 to the Gronigen Team’s K190015 pArs site using the Endy Lab’s expression vector J61002. We loaded J33201 into a kanamycin resistant backbone pSB1K3 and exchanged it with K190015 by perfoming an upstream cut of both to leave the RFP in the J61002 backbone. (Ratio of J33201 to K190015 plasmid DNA was 3:1) | ||
If the increased expression of ArsR had the effect of increasing the threshold of the arsenic response any resulting colonies of J33201 in J61002 we predicted would appear white. | If the increased expression of ArsR had the effect of increasing the threshold of the arsenic response any resulting colonies of J33201 in J61002 we predicted would appear white. | ||
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Not wanting apparent success to get in our way we hypothesized that the addition of a second ArsR gene under pArs control should further raise the threshold of the device. We therefore added a terminiator to J33201 and coupled it upstream from J33201 and as a control K190015. A down stream cut of K190015 from J661002 expression plasmid yields pArs+RBS+RFP+Term. | Not wanting apparent success to get in our way we hypothesized that the addition of a second ArsR gene under pArs control should further raise the threshold of the device. We therefore added a terminiator to J33201 and coupled it upstream from J33201 and as a control K190015. A down stream cut of K190015 from J661002 expression plasmid yields pArs+RBS+RFP+Term. | ||
| - | = | + | <img src=http://fc08.deviantart.net/fs10/i/2006/095/d/c/The_Water_by_leepawlowicz.jpg |
| - | + | <b> Make sure your water is the best, use our arsenic test!!! </b> | |
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Revision as of 02:18, 4 October 2012
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Overall project
Project Details
We compared the sensitivity of J33201 to the Gronigen Team’s K190015 pArs site using the Endy Lab’s expression vector J61002. We loaded J33201 into a kanamycin resistant backbone pSB1K3 and exchanged it with K190015 by perfoming an upstream cut of both to leave the RFP in the J61002 backbone. (Ratio of J33201 to K190015 plasmid DNA was 3:1) If the increased expression of ArsR had the effect of increasing the threshold of the arsenic response any resulting colonies of J33201 in J61002 we predicted would appear white.
Not wanting apparent success to get in our way we hypothesized that the addition of a second ArsR gene under pArs control should further raise the threshold of the device. We therefore added a terminiator to J33201 and coupled it upstream from J33201 and as a control K190015. A down stream cut of K190015 from J661002 expression plasmid yields pArs+RBS+RFP+Term.
<img src=http://fc08.deviantart.net/fs10/i/2006/095/d/c/The_Water_by_leepawlowicz.jpg Make sure your water is the best, use our arsenic test!!!
