Part:BBa_K346004

RBS(B0034)_MBP(lead metal binding peptide egineered from PbrR)+Terminator(B0015)

RBS+mbp(codes for the lead binding peptide)+terminator.

This part was designed to function as lead binding peptide in our project.It is expressed in the cytosol and when fused with the periplasm protein Dsba and membrane protein lpp-ompa, it will be expressed and translocated to the periplasm and membrane.In short, this part function as the fundamental metal binding peptide of lead in our project.

Description

MerR family TFs share a high similarity at the C-terminal metal binding domain , which indicates a similar metal recognition mechanism and metal-protein complex structure.Previous work shows that C-terminal metal binding domain can act as a metal accumulator without the help of the N-terminal DNA binding domain.Function test of mercury MBP showed that our whole-cell bioabsorbents can absorb more than 50% of 10-6 M Hg (II) in 120 minutes, indicating that cells expressing certain metal binding peptide engineered via our method can decontaminate corresponding metal ion from aquatic environment. This design has been successfully applied to another homolog of MerR family – PbrR, the lead responsive regulator.In our project, we try to tandem two metal binding domains together to make a high performance and less energy consuming metal binding peptide. Our bacteria achieved an equivalent lead accumulation capacity as the mercury MBP, which proved validness of our engineering strategy. This part works as the core part in the lead bioabsorption device.

The figure shows the structure of PbrR and MBP(lead). The left figure is PbrR a member of MerR family and the right figure shows the predicted structure of resulted metal binding peptide of lead, which is engineered from PbrR. lead ions are indicated as black balls in metal binding pockets.

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]