Part:BBa_K3332048
J23100-RBS-Lpp-OmpA-iNAP-terminator
iNAP is fused at N-terminal with Lpp-OmpA anchoring protein. We use K880005 to construct the expression system and anchor iNAP on the surface of E.coli.
Biology
Lpp-OmpA is an anchor protein from E. Coli, which can anchor its passenger protein to the cell membrane. It is constituted of 1-9 amino acids of Lpp and 46-159 amino acids of OmpA. It 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 N terminal with Lpp-OmpA 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 use BBa_K880005 to construct the expression system and obtain the composite part BBa_K3332048, which may achieve surface display of iNap on our engineered 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 Lpp-OmpA-iNap
Reference
- ↑ 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 798
Illegal XhoI site found at 824 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 564
- 1000COMPATIBLE WITH RFC[1000]
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