Difference between revisions of "Part:BBa K5375000"
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K5375000 SequenceAndFeatures</partinfo>
 
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===Functional Parameters===
 
===Functional Parameters===
 
<partinfo>BBa_K5375000 parameters</partinfo>
 
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<span id="origin"></span>
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= Origin =
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Synthesized by company
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<span id="properties"></span>
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= Properties =
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Expression of Profilin 3
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<span id="usage-and-biology"></span>
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= Usage and Biology =
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Profilin is a ubiquitously expressed protein that serves as a crucial regulator of actin polymerization. The cytoskeleton plays an integral role in various cellular physiological processes including locomotion, endocytosis, metabolism, signal transduction, and gene transcription. Pollen from trees, herbaceous plants, and weeds has been implicated in allergic reactions. To date, eleven specific IgE-reactive allergens derived from herbaceous plant pollen have been identified. Research indicates that these pollen allergens are categorized into several protein families such as expansins, profilins, and calcium-binding proteins. Profilins function as pan-allergens; they are conserved across plant species and can elicit allergic responses in multiple organisms.
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<span id="cultivation-purification-sds-page"></span>
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= Cultivation, Purification and SDS-PAGE =
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We constructed pPICZαA-Profilin3-sfGFP and pPICZαA-HSP70-sfGFP using homologous recombination. The Profilin 3-sfGFP sequence was amplified by PCR with a length of 1146 bp. The Figure 1 indicates the band consistent with the results. For linearization of the pPICZαA plasmid, the EcoRI and SalI digestion and gel recovery, the pPICZαA fragment was obtained. The Figure 2 shows a band consistent with the target size. It indicates successful linearization of the pPICZαA plasmid.
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<div style="text-align:center;">
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    <img src="https://static.igem.wiki/teams/5375/1.png" width="70%" style="display:block; margin:auto;"
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        alt="Target gene PCR amplification diagram" >
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    <div style="text-align:center;">
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        <caption>
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        Figure 1. Target gene PCR amplification diagram.
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        </caption>
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    </div>
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</div>
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</html>
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<span id="measurement-characterization"></span>
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= Measurement and Characterization =
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We utilized DNA sequencing to determine the full nucleotide sequence of our reconstructed plasmids.
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<html>
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<div style="text-align:center;">
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    <img src="https://static.igem.wiki/teams/5375/2.png" width="70%" style="display:block; margin:auto;"
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        alt="Sanger sequencing of constructed plasmid" >
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    <div style="text-align:center;">
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        <caption>
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        Figure 2. Sanger sequencing of constructed plasmid.
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        </caption>
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    </div>
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</div>
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</html>
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<span id="reference"></span>
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= Reference =
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Davey R. J., & Moens P. D. (2020). Profilin: many facets of a small protein. *Biophysical reviews*, 12(4), 827–849. https://doi.org/10.1007/s12551-020-00723-3
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Chen M., Xu J., Devis D., Shi J., Ren K., Searle I., & Zhang D. (2016). Origin and Functional Prediction of Pollen Allergens in Plants. *Plant physiology*, 172(1), 341–357. https://doi.org/10.1104/pp.16.00625

Revision as of 08:30, 25 September 2024

Profilin 3

Profilin 3


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 172
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Origin

Synthesized by company

Properties

Expression of Profilin 3

Usage and Biology

Profilin is a ubiquitously expressed protein that serves as a crucial regulator of actin polymerization. The cytoskeleton plays an integral role in various cellular physiological processes including locomotion, endocytosis, metabolism, signal transduction, and gene transcription. Pollen from trees, herbaceous plants, and weeds has been implicated in allergic reactions. To date, eleven specific IgE-reactive allergens derived from herbaceous plant pollen have been identified. Research indicates that these pollen allergens are categorized into several protein families such as expansins, profilins, and calcium-binding proteins. Profilins function as pan-allergens; they are conserved across plant species and can elicit allergic responses in multiple organisms.

Cultivation, Purification and SDS-PAGE

We constructed pPICZαA-Profilin3-sfGFP and pPICZαA-HSP70-sfGFP using homologous recombination. The Profilin 3-sfGFP sequence was amplified by PCR with a length of 1146 bp. The Figure 1 indicates the band consistent with the results. For linearization of the pPICZαA plasmid, the EcoRI and SalI digestion and gel recovery, the pPICZαA fragment was obtained. The Figure 2 shows a band consistent with the target size. It indicates successful linearization of the pPICZαA plasmid.

Target gene PCR amplification diagram
Figure 1. Target gene PCR amplification diagram.

Measurement and Characterization

We utilized DNA sequencing to determine the full nucleotide sequence of our reconstructed plasmids.

Sanger sequencing of constructed plasmid
Figure 2. Sanger sequencing of constructed plasmid.

Reference

Davey R. J., & Moens P. D. (2020). Profilin: many facets of a small protein. *Biophysical reviews*, 12(4), 827–849. https://doi.org/10.1007/s12551-020-00723-3

Chen M., Xu J., Devis D., Shi J., Ren K., Searle I., & Zhang D. (2016). Origin and Functional Prediction of Pollen Allergens in Plants. *Plant physiology*, 172(1), 341–357. https://doi.org/10.1104/pp.16.00625