Part:BBa_K3332050
J23100-RBS-iNAP-BrkA-terminator
iNAP is fused at C-terminal with BrkA anchoring protein. We use K880005 to construct the expression system and anchor iNAP on the surface of E.coli.
Biology
BrkA is an anchor protein from Bordetella pertussis having β-barrel structure. It can anchor its passenger protein to the cell membrane and has been widely used in cell-surface display. iNap is a chimera of circularly permuted YFP (cpYFP) and the NADP(H) binding domain of Rex from Thermus aquaticus (T-Rex). Upon NADPH binding, iNap shows apparent fluorescence changes. iNap is fused at C terminal with BrkA so that iNap can be displayed on the surface of E. coli.[1][2]
- Fig 1. Mechanism of iNap on the surface of E. coli
Usage
Here, we used BBa_K880005 to construct the expression system and obtained the composite part BBa_K3332050, which may achieve surface display of iNap on our engineering bacteria. Due to the limited time, we did not get the gene in time. As a result, there is a lack of data about this part. The progress of this part remains in the stage of design.
- Fig 2. Gene circuit of iNap-BrkA
References
- ↑ Zou Y, Wang A, Shi M, et al. Analysis of redox landscapes and dynamics in living cells and in vivo using genetically encoded fluorescent sensors[J]. Nat Protoc, 2018, 13(10): 2362-2386.
- ↑ http://2016.igem.org/Team:TJUSLS_China
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 333
Illegal XhoI site found at 359 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 99
Illegal NgoMIV site found at 1520
Illegal NgoMIV site found at 1949
Illegal NgoMIV site found at 2585 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 2553
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