Part:BBa_K4385020

Zinc-inducible surface display

This part is composed of ZraSR two-component signal transport system, PzraP and OmpC-ZBP.

Usage and Biology

When the zinc ion concentration in the environment exceeds a certain concentration, the PzraP promoter promotes the transcription of downstream OmpC-ZBP, and realizes the adsorption of zinc ions in the environment. Add 6*Hig Tag to this sequence to purify recombinant protein.

Characterization

PZraP-OmpC-ZBP-ST was then constructed so that the OmpC-ZBP would be expressed only in the presence of zinc ions in the environment (Fig. 1A). The enzyme digestion and agarose gel electrophoresis were performed to verify the construction of the PZraP-OmpC-ZBP-ST (Fig. 1B).

After 4 hrs of induction with 1, 2 mM ZnSO4 respectively, the concentration of zinc ions in the supernatant was also measured by Micro Serum Zinc Concentration Assay Kit. The result also shows that our engineered bacteria have better capacity of zinc absorption (Fig. 1C).

Figure 1.Validation of the absorption of zinc ions with PZraP.(A) Digestion and gel electrophoresis of PzraP-OmpC-ZBP-ST (B) The measurement of the concentration of zinc ions in the supernatant. Zn2+ was added when the OD600 value of bacteria reached 0.5. The concentration in supernatant was measured after 4 hrs of induction and normalized by the same OD600 value. *，P<0.05 from control group.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 616
  Illegal SpeI site found at 775
  Illegal PstI site found at 1123
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 616
  Illegal NheI site found at 1506
  Illegal NheI site found at 1529
  Illegal SpeI site found at 775
  Illegal PstI site found at 1123
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 616
  Illegal BglII site found at 324
  Illegal BglII site found at 1641
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 616
  Illegal SpeI site found at 775
  Illegal PstI site found at 1123
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 616
  Illegal SpeI site found at 775
  Illegal PstI site found at 1123
  Illegal AgeI site found at 442
  
• 1000
  COMPATIBLE WITH RFC[1000]