Difference between revisions of "Part:BBa K174015"
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This cadmium sensor is built with binding sites for two metal sensor proteins, ArsR and CzrA (1,2). They both work as repressors and they are relieved from binding to the DNA when they are bound to various metals. ArsR can bind to cadmium, silver, copper and arsenic whereas CzrA can bind to zinc, cobalt, nickel and cadmium. Cadmium can bind to both proteins and using a combinatorial approach enables us to filter out sensing other metals rather than cadmium. | This cadmium sensor is built with binding sites for two metal sensor proteins, ArsR and CzrA (1,2). They both work as repressors and they are relieved from binding to the DNA when they are bound to various metals. ArsR can bind to cadmium, silver, copper and arsenic whereas CzrA can bind to zinc, cobalt, nickel and cadmium. Cadmium can bind to both proteins and using a combinatorial approach enables us to filter out sensing other metals rather than cadmium. | ||
| − | When cadmium is present both proteins are unbound from this AND gate and the promoter region controlled by the repressors becomes free to drive the expression of downstream genes. | + | When cadmium is present both proteins are unbound from this AND gate and the promoter region controlled by the repressors becomes free to drive the expression of downstream genes such as Gfp as an indicator or other transcription factors. |
| + | To create our device we combined ArsR binding site with ''cadA'' promoter from ''B. subtilis''. CadA works as an efflux system to take the metals out in the cell. It has a binding site for CzrA just after the promoter region. | ||
| + | |||
For more information about this part, go to Newcastle iGEM 2009 [http://2009.igem.org/Team:Newcastle/Project/Overview Overview] and [http://2009.igem.org/Team:Newcastle/Metalsensing Metal Sensing] pages. | For more information about this part, go to Newcastle iGEM 2009 [http://2009.igem.org/Team:Newcastle/Project/Overview Overview] and [http://2009.igem.org/Team:Newcastle/Metalsensing Metal Sensing] pages. | ||
Revision as of 22:27, 20 October 2009
Cadmium Sensor
This cadmium sensor is built with binding sites for two metal sensor proteins, ArsR and CzrA (1,2). They both work as repressors and they are relieved from binding to the DNA when they are bound to various metals. ArsR can bind to cadmium, silver, copper and arsenic whereas CzrA can bind to zinc, cobalt, nickel and cadmium. Cadmium can bind to both proteins and using a combinatorial approach enables us to filter out sensing other metals rather than cadmium.
When cadmium is present both proteins are unbound from this AND gate and the promoter region controlled by the repressors becomes free to drive the expression of downstream genes such as Gfp as an indicator or other transcription factors.
To create our device we combined ArsR binding site with cadA promoter from B. subtilis. CadA works as an efflux system to take the metals out in the cell. It has a binding site for CzrA just after the promoter region.
For more information about this part, go to Newcastle iGEM 2009 [http://2009.igem.org/Team:Newcastle/Project/Overview Overview] and [http://2009.igem.org/Team:Newcastle/Metalsensing Metal Sensing] pages.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 200
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
References
- Moore, C. M. and J. D. Helmann (2005). "Metal ion homeostasis in Bacillus subtilis." Current Opinion in Microbiology 8(2): 188-195.
- Harvie, D.R., et al., Predicting metals sensed by ArsR-SmtB repressors: Allosteric interference by a non-effector metal. Molecular Microbiology, 2006. 59(4): p. 1341-1356.