Part:BBa_K3619021

Fusion Protein GST-Ap24, Antimicrobial after Enterokinase cleavage

A fusion protein of 6xHis – Glutathione-s-transferase - Enterokinase cleavage site - Ap24.

A 6xhistidine affinity tag allows for later purification of protein, only prior to protease cleavage.

Ap24 encodes a antimicrobial peptide (AMP) which can be further read about in part BBa_K3619005. The Glutathione-s-transferase fusion partner is meant to increase expression rates, solubility and aid in correct folding of the AMP (see BBa_K3619002). Antimicrobial activity is also likely decreased until the protein is cleaved with enterokinase (see BBa_K3619003). Decrease in antimicrobial activity should prevent it from killing or inhibiting the expressing microbe.

This part was created by the iGEM LUND 2020 team based on previous effective AMP production as fusion partners, which can be seen in [1].

1. https://pubmed.ncbi.nlm.nih.gov/28807840/

The part is designed and codon optimized for expression in E. coli K12.

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 NgoMIV site found at 177
  
• 1000
  COMPATIBLE WITH RFC[1000]