Team:Kyoto/Contribution
Contribution
-This year, our team could make an innovative contribution to future iGEM, mainly through adding the new parts to the iGEM registry.
First, the four parts listed below are used for the peptide production. These parts are the sequences for the new purification method, ELK16 system. This system had not been adopted by other past iGEM teams.
As you can see the below parts, they are composed of ELK16 that has the ability to self-assemble, Mxe GyrA intein that is capable of self-disconnecting by DTT, PT linker, and the targeted protein. In this system, when the fused protein is produced, ELK16, which is the part of it, self-aggregates and precipitates each other. After that, taking this aggregate and adding DTT in order to cut the N terminus of the intein, we can get the targeted protein. Compared to the other methods, the expensive column is not needed, and the additional process to separate the tag from the protein of interest could be omitted.
-Although we could make sure that only one kind of peptide could be synthesized by this method, if we could improve it based on this failure, this system should be one of the most efficient purification methods.
（ELK16systemのPartsを載せる）
Part① Part② Part③ Part④ （Part⑤？）
In addition to the above parts, we also made them that are composed of His tag, Mxe GyrA intein, PT linker, and the targeted protein. In the ELK16 system, in order to recover the targeted protein, the fused protein needs to be precipitated. Some proteins could not solubilize again once they have formed the aggregates. In this case, these parts might be useful because they have His tag instead of ELK16.
This purification system with His tag and intein has contributed greatly to the development of recombinant protein production and purification because it might save time removing tag. The intein that we used this time is cut only when adding DTT. Therefore, they should be useful to improve the protein purification for future iGEM teams. In addition to this, the intein is also used for tag-free purification currently being actively researched, so these parts would be applied in a variety of ways.
（HistagとインテインのPartsを載せる）
Part⑥ Part⑦ Parts⑧ Parts⑨ Parts⑩
Reference
https://link.springer.com/article/10.1007/s00449-020-02382-w
https://www.sciencedirect.com/science/article/pii/S0076687914000676?via%3Dihub
https://microbialcellfactories.biomedcentral.com/articles/10.1186/1475-2859-4-32
First, the four parts listed below are used for the peptide production. These parts are the sequences for the new purification method, ELK16 system. This system had not been adopted by other past iGEM teams.
As you can see the below parts, they are composed of ELK16 that has the ability to self-assemble, Mxe GyrA intein that is capable of self-disconnecting by DTT, PT linker, and the targeted protein. In this system, when the fused protein is produced, ELK16, which is the part of it, self-aggregates and precipitates each other. After that, taking this aggregate and adding DTT in order to cut the N terminus of the intein, we can get the targeted protein. Compared to the other methods, the expensive column is not needed, and the additional process to separate the tag from the protein of interest could be omitted.
-Although we could make sure that only one kind of peptide could be synthesized by this method, if we could improve it based on this failure, this system should be one of the most efficient purification methods.
（ELK16systemのPartsを載せる）
Part① Part② Part③ Part④ （Part⑤？）
In addition to the above parts, we also made them that are composed of His tag, Mxe GyrA intein, PT linker, and the targeted protein. In the ELK16 system, in order to recover the targeted protein, the fused protein needs to be precipitated. Some proteins could not solubilize again once they have formed the aggregates. In this case, these parts might be useful because they have His tag instead of ELK16.
This purification system with His tag and intein has contributed greatly to the development of recombinant protein production and purification because it might save time removing tag. The intein that we used this time is cut only when adding DTT. Therefore, they should be useful to improve the protein purification for future iGEM teams. In addition to this, the intein is also used for tag-free purification currently being actively researched, so these parts would be applied in a variety of ways.
（HistagとインテインのPartsを載せる）
Part⑥ Part⑦ Parts⑧ Parts⑨ Parts⑩
Reference
https://link.springer.com/article/10.1007/s00449-020-02382-w
https://www.sciencedirect.com/science/article/pii/S0076687914000676?via%3Dihub
https://microbialcellfactories.biomedcentral.com/articles/10.1186/1475-2859-4-32