Difference between revisions of "Part:BBa M50006:Design"

Revision as of 06:24, 12 December 2016

zraP promoter for 2-component lead sensing system in E. coli

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE 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, 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.

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. 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

@@ Line 11: / Line 11: @@
 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.
+[[File:ZraS System NDM.png]]
+Our E. coli biosensor is built to function based on the ZraS/ZraR system, 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.
@@ Line 17: / Line 19: @@
 [[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. 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===