Difference between revisions of "Part:BBa M36921"

 
 
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<partinfo>BBa_M36921 short</partinfo>
 
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This composite part contains a promoter and ribosome-binding-site, a gene for ZntR, a terminator, and a promoter for ZntA. In the absence of Zinc, the ZntR protein binds to the ZntA binding site. In the presence of zinc, ZntR undergoes a conformation, does not attach to the binding site, and allows for transcription of ZntA. Essentially, this part acts as a sensor for the presence of zinc.
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This composite part contains a constitutive promoter, ribosome-binding-site, a gene for ZntR, an E. Coli terminator, and a promoter that is ZntR regulated. In the absence of Zinc ions, the ZntR protein binds to the ZntR promoter binding site. In the presence of zinc ions, ZntR undergoes a conformation, does not attach to the binding site, and allows for transcription of our actuator.  
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===Usage and Biology===
 
===Usage and Biology===
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We transfected our plasmid into an ''E. coli'' cell line and wanted to employ it as a primary screening technique for optimum concentrations of Zn2+ in soil. The promoter and ZntR regulatory gene of the znt operon are naturally existing in E. coli and were first sequenced and amplified from E. coli DH5a genomic DNA.
  
 
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===Functional Parameters===
 
===Functional Parameters===
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This system will be placed into a Comet plasmid with a high-copy origin of replication and Kanamycin resistance as its selection marker. We observed that an increasing concentration of Zn2+ did correlate with an increasing amount of Comet fluorescence. However, due to a comparison with controls and further statistical analysis, the composite Zn sensor produced unspecific effects in relation to Zn2+ concentrations.
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<partinfo>BBa_M36921 parameters</partinfo>
 
<partinfo>BBa_M36921 parameters</partinfo>
 
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Latest revision as of 06:43, 5 December 2014

Zinc Sensor

This composite part contains a constitutive promoter, ribosome-binding-site, a gene for ZntR, an E. Coli terminator, and a promoter that is ZntR regulated. In the absence of Zinc ions, the ZntR protein binds to the ZntR promoter binding site. In the presence of zinc ions, ZntR undergoes a conformation, does not attach to the binding site, and allows for transcription of our actuator.


Usage and Biology

We transfected our plasmid into an E. coli cell line and wanted to employ it as a primary screening technique for optimum concentrations of Zn2+ in soil. The promoter and ZntR regulatory gene of the znt operon are naturally existing in E. coli and were first sequenced and amplified from E. coli DH5a genomic DNA.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 194
  • 12
    INCOMPATIBLE WITH RFC[12]
    Unknown
  • 21
    INCOMPATIBLE WITH RFC[21]
    Unknown
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 194
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 194
  • 1000
    COMPATIBLE WITH RFC[1000]


Functional Parameters

This system will be placed into a Comet plasmid with a high-copy origin of replication and Kanamycin resistance as its selection marker. We observed that an increasing concentration of Zn2+ did correlate with an increasing amount of Comet fluorescence. However, due to a comparison with controls and further statistical analysis, the composite Zn sensor produced unspecific effects in relation to Zn2+ concentrations.


emissionComet Fluorescence (wavelength 403 nm : 511 nm)
resistanceKanamycin