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]]
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This part is for new purification method, ELK16 system.
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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>
  
  
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Binding the sequence of targeted protein to the N-terminus of intein, we can produce the peptide with ELK16 method.  
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Fusing the sequence of targeted protein to the N-terminus of intein, we can purify the peptide with ELK16 method.  
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In our experiment, <partinfo>BBa_K3815002</partinfo> could be successfully purified in ELK16.
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In our experiment, <partinfo>BBa_K3815002</partinfo> could be successfully purified with ELK16 system.
  
 
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Latest revision as of 01:53, 22 October 2021


Mxe GryA intein-PT-linker-ELK16 for peptide production

Fig1.The mechanism of ELK16

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


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 13
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 481
  • 1000
    COMPATIBLE 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.