Part:BBa_K5302017

pBBR-OmpA-8IIU

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. Colony PCR results of pBBR-OmpA-8IIU

Figure 2. SDS-PAGE analysis of pBBR1MCS-INP-miniZ expression in Escherichia coli Nissle 1917

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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 3. 8IIU chainA features, from genbank

Figure 4. 8IIU chainB features, from genbank

Figure 5. 8IIU structure, from genbank

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 3931
  Illegal EcoRI site found at 4301
  Illegal XbaI site found at 3140
  Illegal PstI site found at 1967
  Illegal PstI site found at 3128
  Illegal PstI site found at 4033
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 3931
  Illegal EcoRI site found at 4301
  Illegal PstI site found at 1967
  Illegal PstI site found at 3128
  Illegal PstI site found at 4033
  Illegal NotI site found at 1008
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 3931
  Illegal EcoRI site found at 4301
  Illegal BglII site found at 1754
  Illegal BamHI site found at 3146
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 3931
  Illegal EcoRI site found at 4301
  Illegal XbaI site found at 3140
  Illegal PstI site found at 1967
  Illegal PstI site found at 3128
  Illegal PstI site found at 4033
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 3931
  Illegal EcoRI site found at 4301
  Illegal XbaI site found at 3140
  Illegal PstI site found at 1967
  Illegal PstI site found at 3128
  Illegal PstI site found at 4033
  Illegal NgoMIV site found at 2418
  Illegal NgoMIV site found at 2701
  Illegal NgoMIV site found at 4959
  Illegal AgeI site found at 4799
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 1330
  Illegal SapI.rc site found at 2267
  Illegal SapI.rc site found at 2477