Difference between revisions of "Part:BBa K3815000"
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<partinfo>BBa_K3815000 short</partinfo> | <partinfo>BBa_K3815000 short</partinfo> | ||
[[File:ELK16.png|300px|right|thumb|Fig1.The mechanism of ELK16]] | [[File:ELK16.png|300px|right|thumb|Fig1.The mechanism of ELK16]] | ||
| − | This part is for new purification method, ELK16 system. | + | This part is for a new purification method, the ELK16 system. |
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. In this system, we do not have to need the expensive column or could save time removing tags.<br> | 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. In this system, we do not have to need the expensive column or could save time removing tags.<br> | ||
Latest revision as of 01:53, 22 October 2021
Mxe GryA intein-PT-linker-ELK16 for peptide production
This part is for a new purification method, the ELK16 system.
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. In this system, we do not have to need the expensive column or could save time removing tags.
Fusing the sequence of targeted protein to the N-terminus of intein, we can purify the peptide with ELK16 method.
In our experiment, BBa_K3815002 could be successfully purified with ELK16 system.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 13
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 481
- 1000COMPATIBLE WITH RFC[1000]
Reference
1Wang, M., Zheng, K., Lin, J., Huang, M., Ma, Y., Li, S., Luo, X., and Wang, J. (2018). Rapid and efficient production of cecropin A antibacterial peptide in Escherichia coli by fusion with a self-aggregating protein. BMC Biotechnol. 18, 62.