Part:BBa_K3332005
iNAP-BrkA
iNAP is fused at C-terminal with BrkA anchoring protein. We use BBa_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]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 272
Illegal XhoI site found at 298 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 38
Illegal NgoMIV site found at 1459
Illegal NgoMIV site found at 1888
Illegal NgoMIV site found at 2524 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 2492
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