Part:BBa_K3815005
NOP1v-Mxe GryA intein-PT-linker-ELK16
Description of this part
Targeted protein
This part is for the purification of NOP1v.This is made by adding a valine to the N-terminus of NOP1.The effect of it is the same as NOP1. We could not get NOP1 sufficiently when using BBa_K3815004. Then, we thought that considering N-end-rule, the N-terminus of NOP1 might have a negative effect on the recovery of peptide. Therefore, this part is made.
Sequence and Features
Purification system
In order to purify the peptide, we adopted ELK16 system that the past iGEM teams had not used. This part is composed of ELK16, Mxe GyrA intein, and PT linker. When this fused protein is produced, ELK16 self-assembles and precipitates. After that, taking this aggregate and adding DTT to it, the N terminal of Mxe GyrA intein is cut. Then, we can get the targeted protein.
We designed this part, however, we did not actually produce it with this part. We made this peptide with BBa_K3815010.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 120
- 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 120
- 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 120
- 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 120
- 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 120
Illegal NgoMIV site found at 553 - 1000COMPATIBLE WITH RFC[1000]
Reference
1. Hoppen, C., Müller, L., Albrecht, A.C., and Groth, G. (2019). The NOP-1 peptide derived from the central regulator of ethylene signaling EIN2 delays floral senescence in cut flowers. Sci. Rep. 9, 1287
2.Kessenbrock, M., Klein, S.M., Müller, L., Hunsche, M., Noga, G., and Groth, G. (2017). Novel Protein-Protein Inhibitor Based Approach to Control Plant Ethylene Responses: Synthetic Peptides for Ripening Control. Front. Plant Sci. 8, 1528.
3.Tobias, J.W., Shrader, T.E., Rocap, G., and Varshavsky, A. (1991). The N-end rule in bacteria. Science 254, 1374–1377.
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