Part:BBa_K2959008

Expressible Wheat antimicrobial peptide 1b with Erv1p

Since WAMP1b is a peptide with five disulfide bonds, a post-traductional modification hard to replicate on prokaryotic expression systems, the construct allows for the co-expression of the peptide with a truncated version of Erv1p. This is a protein from Saccharomyces cerevisiae capable of catalyzing the formation of disulfide bonds.

This composite part consists of a lacI regulated promoter, ribosome binding site, a coding sequence for WAMP1b as a fusion protein with a 6x His-Tag, another ribosome binding site, the coding sequence for Erv1p, and a double terminator. This construct allows the expression of WAMP1b, an antifungal peptide from Triticum kiharae seeds, in E. coli, as well as the protein Erv1p. Expression can be positively regulated by the addition of IPTG or lactose thanks to the lacl regulated promoter. The part is designed to code for a fusion protein of WAMP1b with a polyhistidine tag (6x His-Tag) at its N-terminus for purification by immobilized metal affinity chromatography.

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 472
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 261