Difference between revisions of "Part:BBa K5317005"
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__NOTOC__ | __NOTOC__ | ||
<partinfo>BBa_K5317005 short</partinfo> | <partinfo>BBa_K5317005 short</partinfo> | ||
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| + | ===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 for cell-based metal detection. | 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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To achieve a cell-based heavy metal sensor, our group generated multiple synthetic MTF1 responsive promoter constructs and tested them in regard to their efficiency. Since Wang and colleagues (2004) suggested a high affinity of MTF-1 towards MREd we synthesized a promoter sequence containing four MREd elements at the positioning of the MREs of the MREwt promoter (reference) to exclude possible disruption of the MTF-1 and MRE interaction. | To achieve a cell-based heavy metal sensor, our group generated multiple synthetic MTF1 responsive promoter constructs and tested them in regard to their efficiency. Since Wang and colleagues (2004) suggested a high affinity of MTF-1 towards MREd we synthesized a promoter sequence containing four MREd elements at the positioning of the MREs of the MREwt promoter (reference) to exclude possible disruption of the MTF-1 and MRE interaction. | ||
| + | =Cloning= | ||
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| − | + | ===Theoretical Part Design=== | |
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| + | The sequence of the MREd site was obtained from Seguin et al., 1987, as well as the positioning and spacer sequences (Searle et al., 1985). The construct was synthesized. | ||
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| + | ===Sequence and Features=== | ||
<partinfo>BBa_K5317005 SequenceAndFeatures</partinfo> | <partinfo>BBa_K5317005 SequenceAndFeatures</partinfo> | ||
Revision as of 08:58, 13 September 2024
4xMREd 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 for cell-based metal detection.
To achieve a cell-based heavy metal sensor, our group generated multiple synthetic MTF1 responsive promoter constructs and tested them in regard to their efficiency. Since Wang and colleagues (2004) suggested a high affinity of MTF-1 towards MREd we synthesized a promoter sequence containing four MREd elements at the positioning of the MREs of the MREwt promoter (reference) to exclude possible disruption of the MTF-1 and MRE interaction.
Cloning
Theoretical Part Design
The sequence of the MREd site was obtained from Seguin et al., 1987, as well as the positioning and spacer sequences (Searle et al., 1985). The construct was synthesized.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]