Part:BBa_K5302020

pBBR-OmpA-Z3C

Just like miniZ, Z3C is also 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 pBBR1MCS-2 plasmid as a backbone and transfered Z3C into Escherichia coli Nissle 1917, and finally succeeded in expressing Z3C.

Figure 1. Colony PCR results of pBBR-OmpA-Z3C

Figure 2. SDS-PAGE analysis of pBBR1MCS-OmpA-Z3C expression in Escherichia coli Nissle 1917 （1）

Figure 3. SDS-PAGE analysis of pBBR1MCS-OmpA-Z3C expression in Escherichia coli Nissle 1917（2）

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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 4. sequence of Z3C and its KD with VEGF

Figure 5. Cartoon representation of the crystal structure of the mini-Z highlighting randomized residues shown as sticks and coloring the helices

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 4100
  Illegal XbaI site found at 3140
  Illegal PstI site found at 1967
  Illegal PstI site found at 3128
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 4100
  Illegal PstI site found at 1967
  Illegal PstI site found at 3128
  Illegal NotI site found at 1008
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 4100
  Illegal BglII site found at 1754
  Illegal BamHI site found at 3146
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 4100
  Illegal XbaI site found at 3140
  Illegal PstI site found at 1967
  Illegal PstI site found at 3128
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 4100
  Illegal XbaI site found at 3140
  Illegal PstI site found at 1967
  Illegal PstI site found at 3128
  Illegal NgoMIV site found at 2418
  Illegal NgoMIV site found at 2701
  Illegal NgoMIV site found at 4758
  Illegal AgeI site found at 3789
  Illegal AgeI site found at 4598
  
• 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