Part:BBa_K4633101

Pveg-AmyE-Based Improved Platform for FimHL and GFP Co-expression in B. subtilis

This composite construct features a Bacillus subtilis constitutive promoter, Pveg (BBa_K143012). Downstream of this promoter, the sequence includes the signal peptide AmyE (BBa_K2273023), a well-established secretion peptide in B. subtilis, followed by a linker (BBa_K2934005), the FimHL domain (BBa_K4633000), a second linker (BBa_K1486037) and GFP (BBa_E0040).

Usage and Biology

This composite part was used by the 2023 Technion team to monitor and validate the expression of their protein of interest, FimHL.

Its main difference, and improvement, from composite part BBa_K4633100, is the addition of a second linker, meant to separate FimHL and GFP so each protein has the proper spatial conformation, and therefore maintains its activity.

FimH, a critical adhesin, is utilized by uropathogenic Escherichia coli (UPEC) to adhere to and subsequently invade epithelial cells within the urinary tract, leading to the onset of urinary tract infections (UTIs). FimH-L specifically represents the lectin domain of FimH, responsible for binding to mannose residues on the cell surface.

The inclusion of GFP serves the purpose of enabling straightforward monitoring of protein expression and secretion throughout the experimental process.

Sequence and Features

Assembly Compatibility:

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

  Illegal BglII site found at 331
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 1395