Difference between revisions of "Part:BBa K3714004"
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− | <center><b>Figure 1 | Improvement of an existing theophylline aptazyme. </b>Cleavage properties of two aptazymes under different theophylline concentration. The aptazymes were transcribed <i>in vitro</i> with certain amount of theophylline, and the products were run a a denatured PAGE. The number above each lane means dilution fold of saturated theophylline aqueous solution at 25℃. | + | <center><b>Figure 1 | Improvement of an existing theophylline aptazyme. </b>Cleavage properties of two aptazymes under different theophylline concentration. The aptazymes were transcribed <i>in vitro</i> with certain amount of theophylline, and the products were run a a denatured PAGE. The number above each lane means dilution fold of saturated theophylline aqueous solution at 25℃. </center> |
To further quantify the outcome of our engineering, we used a lower concentration gradient. Our aptazyme exhibits a 2-fold change of cleavage proportion at ~1mg/ml (0.125x dilution of saturated solution at room temperature) while the origin apatazyme (BBa_J119449) shows only 1.3-fold change (Fig.2). | To further quantify the outcome of our engineering, we used a lower concentration gradient. Our aptazyme exhibits a 2-fold change of cleavage proportion at ~1mg/ml (0.125x dilution of saturated solution at room temperature) while the origin apatazyme (BBa_J119449) shows only 1.3-fold change (Fig.2). | ||
<center>[[File:K3714004-2.jpg]]</center> | <center>[[File:K3714004-2.jpg]]</center> |
Revision as of 23:48, 21 October 2021
Improved theophylline biosensor
An improved aptazyme responsing to theophylline, which means its self-cleaving activity can be inhibited by adding theophylline. This aptazyme shows higher sensibility than an existing part (BBa_J119449).
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
By introducing mutations in loop Ⅰ and loop Ⅱ[1], we achieved an aptazyme with higher responsing fold and less leakage under high theophylline concentration (Fig.1).
To further quantify the outcome of our engineering, we used a lower concentration gradient. Our aptazyme exhibits a 2-fold change of cleavage proportion at ~1mg/ml (0.125x dilution of saturated solution at room temperature) while the origin apatazyme (BBa_J119449) shows only 1.3-fold change (Fig.2).