Difference between revisions of "Part:BBa M36921"
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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. | 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=== | ||
− | 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. | + | 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
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 194
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30
Illegal PstI site found at 194 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 190
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 194
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 194
- 1000COMPATIBLE 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.
emission | Comet Fluorescence (wavelength 403 nm : 511 nm) |
resistance | Kanamycin |