Difference between revisions of "Part:BBa K5459001"

 
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<partinfo>BBa_K5459001 short</partinfo>
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        <p><strong>Regulatory protein for copper detection.</strong><br>This is a part designed for the expression of the CueR protein, utilizing a strong promoter BBa_J23119 to drive its expression for our copper ion detection system. This. part can be used in conjunction with BBa_K5459002, but our experimental results have shown that BBa_K5459002 can also function independently by leveraging the endogenous CueR at the genomic level in E. coli. We have compared the differences between these two scenarios:</p>
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CueR family proteins are key metal regulatory proteins in Escherichia coli that specifically sense monovalent copper ions (Cu&#8314;) in the internal and external cellular environment. CueR belongs to the MerR family of transcriptional regulators and plays a crucial role in regulating copper ion homeostasis and resistance to copper toxicity. When copper ion concentrations rise, CueR binds to copper ions with high affinity. Once copper ions enter the cell, CueR rapidly binds to them, triggering a conformational change in the CueR protein. When used in conjunction with BBa_K5459002, it can detect copper ion concentrations.
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            <img src="https://static.igem.wiki/teams/5459/part-registry1/compration.jpg" alt="图一" style="width: 300px; margin-right: 10px;">
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===Usage and Biology===
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        <p>We have observed that in a two-plasmid system, the use of the BBa_J23119 promoter to initiate CueR expression indeed impacts the fluorescence levels of the entire system. This influence can be harnessed to develop copper ion detectors with varying performance characteristics.<br><strong>High throughput promoter experiment</strong></p>
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            <img src="https://static.igem.wiki/teams/5459/part-registry1/high-throghput1.jpg" alt="图三" style="width: 300px; margin-right: 10px;">
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<span class='h3bb'>Sequence and Features</span>
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            <img src="https://static.igem.wiki/teams/5459/part-registry1/96-ratio-zhuzhuangtu.jpg" alt="图三" style="width: 300px; margin-right: 10px;">
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<partinfo>BBa_K5459001 SequenceAndFeatures</partinfo>
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            <img src="https://static.igem.wiki/teams/5459/part-registry1/96-ratio-heatmap.jpg" alt="图四" style="width: 300px;">
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      <p>We observed that all promoters in the promoter library exhibit a certain level of promoter activity, indicating their significant role in transcriptional regulation. The initiation strength of the promoters shows a degree of variability, with the lowest induction strength being 5-fold, and the highest reaching up to 10-fold. To gain a deeper understanding of the relationship between the activity of these promoters and their sequence characteristics, we employed Sanger sequencing to determine the nucleotide sequences of these promoters. Subsequently, based on the sequencing results, we conducted a correlation analysis between the initiation strength of the promoters and their sequence features, and built a mathematical model accordingly. The detailed construction process and parameter settings of this model can be consulted on our provided "Model" page. Through this model, we aim to reveal the specific mechanisms by which promoter sequence characteristics affect their initiation activity, providing a theoretical foundation and experimental guidance for subsequent research on gene expression regulation.</p>
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===Functional Parameters===
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<partinfo>BBa_K5459001 parameters</partinfo>
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Revision as of 17:08, 27 September 2024

Regulatory protein for copper detection.
This is a part designed for the expression of the CueR protein, utilizing a strong promoter BBa_J23119 to drive its expression for our copper ion detection system. This. part can be used in conjunction with BBa_K5459002, but our experimental results have shown that BBa_K5459002 can also function independently by leveraging the endogenous CueR at the genomic level in E. coli. We have compared the differences between these two scenarios:

图一

We have observed that in a two-plasmid system, the use of the BBa_J23119 promoter to initiate CueR expression indeed impacts the fluorescence levels of the entire system. This influence can be harnessed to develop copper ion detectors with varying performance characteristics.
High throughput promoter experiment

图三 图三 图四

We observed that all promoters in the promoter library exhibit a certain level of promoter activity, indicating their significant role in transcriptional regulation. The initiation strength of the promoters shows a degree of variability, with the lowest induction strength being 5-fold, and the highest reaching up to 10-fold. To gain a deeper understanding of the relationship between the activity of these promoters and their sequence characteristics, we employed Sanger sequencing to determine the nucleotide sequences of these promoters. Subsequently, based on the sequencing results, we conducted a correlation analysis between the initiation strength of the promoters and their sequence features, and built a mathematical model accordingly. The detailed construction process and parameter settings of this model can be consulted on our provided "Model" page. Through this model, we aim to reveal the specific mechanisms by which promoter sequence characteristics affect their initiation activity, providing a theoretical foundation and experimental guidance for subsequent research on gene expression regulation.