Difference between revisions of "Part:BBa K3332003"

Latest revision as of 22:50, 27 October 2020

Lpp-OmpA-iNAP

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

1. ↑ 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
2. ↑ http://2016.igem.org/Team:TJUSLS_China

Sequence and Features