Part:BBa_K4468009

GolS- T7 Terminator- PgolB

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 162
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 2385
  Illegal AgeI site found at 633
  Illegal AgeI site found at 982
  Illegal AgeI site found at 1920
  
• 1000
  COMPATIBLE WITH RFC[1000]

Description

This is a composite component for absorbing Cu²⁺ in solution and activating the promoter PgolB. It consists of GolS- PgolB. As a component, it can provide a powerful aid for plasmid synthesis in molecular cloning.

Usage and Biology

GolS

The GolS of Salmonella is a transcriptional regulator comes from the gold specific family MerR. Its expressed protein GolS plays a role as an operon that responsibles for regulating the promoter PgolB. Upon recognizing and adsorbing extracellular Au³⁺, the expression of downstream genes after promoter PgolB will be activated.
GolS, which has a metal binding loop that can specifically binds Au³⁺ and regulates the expression of genes downstream of PgolB, is the most important part of the regulatory system. Literatures have proved that residues 108-120 of GolS, which are in this metal binding loop, play a decisive role for binding Au³⁺. It was found that if this part of the residues were changed to residues 108-120 of the cognate CueR protein, the new GolS protein not only has Au³⁺ adsorption ability, but also high Cu²⁺ adsorption. CueR and GolS are homologous transcription factors with high sequence similarity and similar metal binding loop. The difference is that CueR specifically binds Cu²⁺ rather than Au³⁺. Considering that the main application occasion of our project is mining wastewater, which tends to contain a large amount of copper element instead of gold element. So we chose the new gols, which replaced the residues of CueR, as our project protein and activate the expression of PgolB by Cu²⁺ induction.

PgolB

The GolS of Salmonella is a transcriptional regulator comes from the gold specific family MerR. Its expressed protein GolS plays a role as an operon that responsibles for regulating the promoter PgolB. Upon recognizing and adsorbing extracellular Au³⁺, the expression of downstream genes after promoter PgolB will be activated.
PgolB is the promoter of the GolS system. The expression of genes after PgolB is strictly regulated by GolS. In the absence of Au³⁺, GolS inhibites PgolB to prevent downstream genes’ expression. Whereas when GolS adsorbs exogenous Au³⁺, the inhibition turns to cease, allowing PgolB to initiate expression.