Part:BBa_K2328025

LPP-OmpA + linker.b + smURFP I

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 750
  
• 1000
  COMPATIBLE WITH RFC[1000]

Usage

This part is a fusion protein of the LPP and smURFP. smURFP (small ultra-red FP) is an far-infrared fluorescent protein. Through this method, we can anchor smURFP on the out membrane of Escherichia coli, then we can use this typical Escherichia coli to connect with biliverdin. And the prokaryote surface display system method is mature enough, we can take use of it to achieve our goals.

Biology

smURFP (small ultra-red FP) is an far-infrared fluorescence. It is desirable for our in vivo imaging because with it molecule less light is scattered, absorbed, or re-emitted by endogenous biomolecules compared with cyan, green, yellow and orange FPs. smURFP can covalently attaches a biliverdin (BV) chromophore without a lyase, and has 642/670 nm excitation - emission peaks, a large extinction coefficient and quantum yield, and photostability comparable to that of eGFP. Surface expression of recombinant proteins was first described more than 30 years ago. Bacterial surface display entails the presentation of recombinant proteins or peptides on the surface of bacterial cells. Escherichia coli is the most frequently used bacterial host for surface display and, as such, a variety of E. coli display systems have been described that primarily promote the surface exposure of peptides and small proteins. And the LPP is a kind of protein that adopt to surface play. So we try this method to display our smURFP on the surface of E.coil so that it can connect the BV added in culture.

Reference

[1] Rodriguez EA,Tran GN , Gross LA, et al. A far-red fluorescent protein evolved from a cyanobacterial phycobiliprotein .[J].NATURE METHODS,2016:763-769.