Part:BBa_K2959005

Expressible Pinus sylvestris Defensin 1 with Erv1p

This composite part consists of a lacI regulated promoter, ribosome binding site, a coding sequence for Pinus sylvestris Defensin 1 as a fusion protein with a 6x His-Tag, and a double terminator. This construct allows the expression of PsDef1, an antifungal peptide from Scots pine seeds, in E. coli. Expression can be positively regulated by the addition of IPTG or lactose thanks to the lacl regulated promoter. The fusion of PsDef1 to a 6x His-Tag is intended for the purification of the peptide by immobilized metal affinity chromatography.

Since PsDef1 is a peptide with four disulfide bonds, a 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.

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 580
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]