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Part:BBa_K4750001
SP+T8 (tpSil3)
This sequence is a fragment of tpSil3 (part BBa_K4750000). It includes the N-terminal 17 amino acid long signal peptide (SP) of tpSil3 and the 37 amino acid long T8 sequence (from residues 164 to 200).
The SP is responsible for co-translational import into the endoplasmic reticulum, and the T8 sequence is sufficient to direct fusion proteins with SP-T8 into the diatom cell wall (N. Poulsen and N. Kröger, unpublished result). (Sheppard et al. 2012) [1]
Use
SP+T8 is designed to be N-terminally fused to a protein of interest and to be consequently expressed in the diatom Thalassiosira pseudonana. In doing so, the fusion protein will be in vivo immobilized into the diatom's cell wall (valve and girdle bands). The underlying method was coined Live Diatom Silica Immobilization (LiDSI) and was developed by the Kröger lab. (Poulsen et al. 2007)[2].
Application
The LiDSI method can be applied in numerous biotechnological fields, such as catalysis, sensing and targeted drug delivery. (Kröger, Dubey, and Kumari 2017)[3]
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 143
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
References
- ↑ Sheppard, V. C., A. Scheffel, N. Poulsen, and N. Kröger. 2012. “Live Diatom Silica Immobilization of Multimeric and Redox-Active Enzymes.” Applied and Environmental Microbiology 78 (1): 211–18. https://doi.org/10.1128/AEM.06698-11.
- ↑ Poulsen, Nicole, Cécile Berne, Jim Spain, and Nils Kröger. 2007. “Silica Immobilization of an Enzyme through Genetic Engineering of the Diatom Thalassiosira Pseudonana.” Angewandte Chemie International Edition 46 (11): 1843–46. https://doi.org/10.1002/anie.200603928.
- ↑ Kröger, Nils, Nidhi Dubey, and Ekta Kumari. 2017. “Immobilization of Proteins on Diatom Biosilica.” In RSC Nanoscience and Nanotechnology. https://doi.org/10.1039/9781788010160-00126.
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