Part:BBa_K5302004

Z3C

This year, the USTC iGEM team has utilized the competitive binding of vascular endothelial growth factor (VEGF) to develop a targeted bacterial therapy for solid tumors. Our quest for the optimal VEGF-binding protein(or peptide) led us to an in-depth exploration of proteins structurally akin to the vascular endothelial growth factor receptor (VEGFR), which we have named VEGFR-like. Z3C is derived from three helix 58-residue Z-domain of staphylococcal protein A, but it has three helix. Z3C also shows great affinity with VEGF(KD=41 nM). We used pBBR plasmid as a backbone and transfered Z3C into Escherichia coli Nissle 1917, and finally succeeded in expressing Z3C.

Figure 1. sequence of Z3C and its KD with VEGF

Figure 2. Cartoon representation of the crystal structure of the mini-Z highlighting randomized residues shown as sticks and coloring the helices

Figure 3. Colony PCR results of pBBR-OmpA-Z3C

Figure 4. SDS-PAGE analysis of pBBR1MCS-OmpA-Z3C expression in Escherichia coli Nissle 1917 (1)

Figure 5. SDS-PAGE analysis of pBBR1MCS-OmpA-Z3C expression in Escherichia coli Nissle 1917 (2)

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