Difference between revisions of "Part:BBa K4468009"

 
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Description
 
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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.
 
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<partinfo>BBa_K4468009 parameters</partinfo>
 
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<h1>'''Description'''</h1>
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This is a composite component for absorbing Cu<sup>2+</sup> 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.
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<h1>'''Usage and Biology'''</h1>
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<h2>'''GolS'''</h2>
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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 Au<sup>3+</sup>, the expression of downstream genes after promoter PgolB will be activated.<br>
 +
GolS, which has a metal binding loop that can specifically binds Au<sup>3+</sup> 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 Au<sup>3+</sup>. 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 Au<sup>3+</sup> adsorption ability, but also high Cu<sup>2+</sup> adsorption. CueR and GolS are homologous transcription factors with high sequence similarity and similar metal binding loop. The difference is that CueR specifically binds Cu<sup>2+</sup> rather than Au<sup>3+</sup>. 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 Cu<sup>2+</sup> induction.
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<h2>'''PgolB'''</h2>
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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 Au<sup>3+</sup>, the expression of downstream genes after promoter PgolB will be activated.<br>
 +
PgolB is the promoter of the GolS system. The expression of genes after PgolB is strictly regulated by GolS. In the absence of Au<sup>3+</sup>, GolS inhibites PgolB to prevent downstream genes’ expression. Whereas when GolS adsorbs exogenous Au<sup>3+</sup>, the inhibition turns to cease, allowing PgolB to initiate expression.

Revision as of 11:07, 30 September 2022


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.