Part:BBa_K5223001
High-intensity 35S promoter
35S promoter is derived from the Cauliflower Mosaic Virus and is commonly used in plant chassis. To improve transcription efficiency, we aim to optimize the 35S promoter. Therefore, we trained a deep learning model using existing data (Jores, Tobias et al., 2021) and achieved promising results, subsequently using this model for promoter optimization. However, due to time and resource constraints, we are currently unable to provide usable experimental data.
Usage and Biology
| Optimized Promoter | |
| Function | High-intensity 35S promoter |
| Use in | Nicotiana Benthamiana |
| Backbone | pZYB11-pEAQ-HT-DEST1 |
| Derived from | Cauliflower Mosaic Virus (CaMV) |
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]
Design and Properties
First, we used TSSPlant to predict the transcription start sites (TSS) corresponding to the wild-type promoter sequences, extracting the -165 to +5 fragments relative to the TSS and performing random single-point mutations on them. The model was then used to predict the strength of the mutants, selecting the ones with the highest strength for the next round of evolution. Ultimately, through 10 and 15 rounds of evolution, we obtained mutants 1 and 2, with predicted strengths of 3.32 times and 2.08 times that of the wild type, respectively.
Figure 2. pET-28a-DspB-C-His plasmid
Discussion and outlook
In the future, we will design experiments to compare the strength differences between the various mutants and the wild type. Additionally, we will conduct more experiments to provide the model with more high-quality data, enhancing the usability of the software we are developing.
Reference
[1] Jores, Tobias et al. “Synthetic promoter designs enabled by a comprehensive analysis of plant core promoters.” Nature plants vol. 7,6 (2021): 842-855. doi:10.1038/s41477-021-00932-y
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