Difference between revisions of "Part:BBa K5317006"
 
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===Usage and Biology===
 
===Usage and Biology===
  
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The MRE-sites containing promoter enables the metal-dependent expression of a downstream positioned reporter gene via the metal ion-dependent transcription factor MTF-1 for cell-based metal detection.  
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The MRE-sites containing promoter enables the metal-dependent expression of a downstream positioned reporter gene via the metal ion-dependent transcription factor MTF-1 (<span class="plainlinks">[https://parts.igem.org/Part:BBa_K5317007 K5317007]</span>) for cell-based metal detection.  
  
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To combine the observations made by Searle and colleagues and Wang and colleagues in 1985 and 2004, respectively, regarding the metal inducibility of a promoter with two MREa sites and a high affinity between MREd and MTF-1, we designed a synthetic promoter with two MREa and two MREd sites that alternate. The aim is possibly increasing the sensitivity and efficiency of the metal-dependent promoter.
+
In order to integrate the findings of Searle and colleagues (1985) and Wang and colleagues (2004) regarding the metal inducibility of a promoter with two MREa sites and a high affinity between MREd and MTF-1, we designed a synthetic promoter with two MREa and two MREd sites that alternate. The aim is to enhance the sensitivity and efficiency of the metal-dependent promoter.
  
 
=Cloning=
 
=Cloning=
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===Theoretical Part Design===
 
===Theoretical Part Design===
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The MREdada promoter sequence was synthesized in order to insert it into the EGFP-C2 backbone (registry entry K3338020) for promoter efficiency testing.  
+
The MREdada promoter sequence was synthesized to insert it into the EGFP-C2 backbone (<span class="plainlinks">[https://parts.igem.org/Part:BBa_K3338020 K3338020]</span>) for promoter efficiency testing.
  
 
===Sequence and Features===
 
===Sequence and Features===
 
<partinfo>BBa_K5317006 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K5317006 SequenceAndFeatures</partinfo>
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 +
=Characterization=
 +
 +
The promoter was analyzed by composing a gene cassette where its placed upstream of the reporter gene EGFP to assess the promoter's functionality and metal-dependent efficiency based on the fluorescent signal. Please visit the <span class="plainlinks">[https://parts.igem.org/Part:BBa_K5317011 K5317011]</span> registry entry to view the results.
  
 
=References=
 
=References=
 +
 +
Searle, P. F., Stuart, G. W., & Palmiter, R. D. (1985). Building a metal-responsive promoter with synthetic regulatory elements. Molecular and cellular biology, 5(6), 1480–1489. https://doi.org/10.1128/mcb.5.6.1480-1489.1985
 +
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Wang, Y., Lorenzi, I., Georgiev, O., & Schaffner, W. (2004). Metal-responsive transcription factor-1 (MTF-1) selects different types of metal response elements at low vs. high zinc concentration. Biological chemistry, 385(7), 623–632. https://doi.org/10.1515/BC.2004.077
  
 
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Latest revision as of 21:34, 1 October 2024


MREdada promoter

Usage and Biology

The MRE-sites containing promoter enables the metal-dependent expression of a downstream positioned reporter gene via the metal ion-dependent transcription factor MTF-1 (K5317007) for cell-based metal detection.

In order to integrate the findings of Searle and colleagues (1985) and Wang and colleagues (2004) regarding the metal inducibility of a promoter with two MREa sites and a high affinity between MREd and MTF-1, we designed a synthetic promoter with two MREa and two MREd sites that alternate. The aim is to enhance the sensitivity and efficiency of the metal-dependent promoter.

Cloning

Theoretical Part Design

The MREdada promoter sequence was synthesized to insert it into the EGFP-C2 backbone (K3338020) for promoter efficiency testing.

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]

Characterization

The promoter was analyzed by composing a gene cassette where its placed upstream of the reporter gene EGFP to assess the promoter's functionality and metal-dependent efficiency based on the fluorescent signal. Please visit the K5317011 registry entry to view the results.

References

Searle, P. F., Stuart, G. W., & Palmiter, R. D. (1985). Building a metal-responsive promoter with synthetic regulatory elements. Molecular and cellular biology, 5(6), 1480–1489. https://doi.org/10.1128/mcb.5.6.1480-1489.1985

Wang, Y., Lorenzi, I., Georgiev, O., & Schaffner, W. (2004). Metal-responsive transcription factor-1 (MTF-1) selects different types of metal response elements at low vs. high zinc concentration. Biological chemistry, 385(7), 623–632. https://doi.org/10.1515/BC.2004.077