Part:BBa_K2922043

Expression system of DsbA-PbBD coding periplasm display of lead binding peptide with T7 promoter

Summary

This part was constructed for high expression of DsbA-PbBD(periplasm display of lead binding peptide). We replaced MBP in BBa_K346030 with PbBD to enhance the lead absorption efficiency.

Figure 1.Schematic diagram of action principle for DsbA-MBP and DsbA-PbBD

Characterization

Inductively coupled plasma mass spectrometry (ICP-MS) was used to accurately test the absorption amount of lead per unit mass of cells. BL21 (DE3) with DsbA-MBP and DsbA-PbBD integration plasmids were separately cultured in medium containing 50 μM Pb(II). The amounts of lead absorbed by the bacteria with different circuits were measured. And as the experimental results shown in Fig.2, the lead absorption efficiency of DsbA-PbBD group was better than that in DsbA-MBP group.

Figure 2.Relative Pb（II）Absorption Capacity Per Unit Mass（DsbA-MBP &DsbA-PbBD）

Reference

1. C. Hui et al., Surface display of PbrR on Escherichia coli and evaluation of the bioavailability of lead associated with engineered cells in mice. Sci Rep 8, 5685 (2018).
2. C. Y. Hui, Y. Guo, X. Q. Yang, W. Zhang, X. Q. Huang, Surface display of metal binding domain derived from PbrR on Escherichia coli specifically increases lead(II) adsorption. Biotechnol Lett 40, 837-845 (2018).

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 317
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 149
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 715