Part:BBa_K5302001

8IIU

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.8IIU is an anti-VEGF nanobody, and it has 2 chains, chainA sequence: mdvqlvesgg glvqpggslr lscaasgrtf ssysmgwfrq apgkerefvv aiskggykyd avslegrfti srdnakntvy lqinslrped tavyycassr aygssrlrla dtyeywgqgt lvtvss. chainB sequence:mdvqlvesgg glvqpggslr lscaasgrtf ssysmgwfrq apgkerefvv aiskggyky avslegrfti srdnakntvy lqinslrped tavyycassr aygssrlrla dtyeywgqgt lvtvss. These two chains all show great affinity with VEGF, we used pBBR plasmid as a backbone and transfered 8IIU into Escherichia coli Nissle 1917, and finally succeeded in expressing 8IIU.

Figure 1. 8IIU chainA features, from genbank

Figure 2. 8IIU chainB features, from genbank

Figure 3. 8IIU structure, from genbank

Figure 4. Colony PCR results of pBBR-OmpA-8IIU

Figure 5. Colony PCR results of pBBR-INP-8IIU (1)

Figure 6. Colony PCR results of pBBR-INP-8IIU (2)

Figure 7. Colony PCR results of pBBR-INP-8IIU (3)

Figure 8. SDS-PAGE analysis of pBBR1MCS-OmpA-8IIU expression in Escherichia coli Nissle 1917

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]