Difference between revisions of "Part:BBa M50006:Design"
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We found the zraP promoter sequence from a 2014 online research paper from Maruthamuthu, Ganesh, Ravikumar, and Hong, and removed the forward and end primer sequences to isolate the promoter sequence. | We found the zraP promoter sequence from a 2014 online research paper from Maruthamuthu, Ganesh, Ravikumar, and Hong, and removed the forward and end primer sequences to isolate the promoter sequence. | ||
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+ | [["File:NanodropTheMicDeviceDesign.png"]] | ||
We also replaced the ribosome binding site (RBS) provided in the sequence with DNA 2.0’s default high-strength RBS. Downstream of the zraP promoter is Comet (GFP), which will be expressed when zraP is activated due to the extracellular concentration of lead(II) ions (Figure 2). A lead-binding peptide, OmpC, on the outer surface of E. coli cells is able to bind lead(II) ions, thus activating the cell response through the ZraS/ZraR two component system. | We also replaced the ribosome binding site (RBS) provided in the sequence with DNA 2.0’s default high-strength RBS. Downstream of the zraP promoter is Comet (GFP), which will be expressed when zraP is activated due to the extracellular concentration of lead(II) ions (Figure 2). A lead-binding peptide, OmpC, on the outer surface of E. coli cells is able to bind lead(II) ions, thus activating the cell response through the ZraS/ZraR two component system. |
Revision as of 06:21, 12 December 2016
zraP promoter for 2-component lead sensing system in E. coli
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
No Bsa1 sites, less than 2,000 base pairs total
E. coli endogenous ZraS/ZraR two-component system
Our E. coli biosensor is built to function based on the ZraS/ZraR system (Figure 1), a two-component endogenous stimulus-sensing apparatus within E. coli.
We found the zraP promoter sequence from a 2014 online research paper from Maruthamuthu, Ganesh, Ravikumar, and Hong, and removed the forward and end primer sequences to isolate the promoter sequence.
"File:NanodropTheMicDeviceDesign.png"
We also replaced the ribosome binding site (RBS) provided in the sequence with DNA 2.0’s default high-strength RBS. Downstream of the zraP promoter is Comet (GFP), which will be expressed when zraP is activated due to the extracellular concentration of lead(II) ions (Figure 2). A lead-binding peptide, OmpC, on the outer surface of E. coli cells is able to bind lead(II) ions, thus activating the cell response through the ZraS/ZraR two component system.
Source
ZraP gene
References
ZraP:Gene. (2013, September 13). Retrieved October 25, 2016, from http://ecoliwiki.net/colipedia/index.php/zraP:Gene
Maruthamuthu, M. K., et al. (2014, November 30). Evaluation of zraP gene expression characteristics and construction of a lead (Pb) sensing and removal system in a recombinant Escherichia coli. Biotechnol Lett, 37(2015), 659-664. doi:DOI 10.1007/s10529-014-1732-x