Difference between revisions of "Part:BBa K3332003"

 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>

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


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 737
    Illegal XhoI site found at 763
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 503
  • 1000
    COMPATIBLE WITH RFC[1000]