Difference between revisions of "Part:BBa K3815010"
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<partinfo>BBa_K3815010 short</partinfo>
 
<partinfo>BBa_K3815010 short</partinfo>
  
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This part is made of Hist tag for the peptide production. After producing peptide with His tag, it is recovered by Ni chromatography. Finally, adding DTT to it, we cut the N terminal of Mxe GyrA intein. Then, we can get the targeted protein.In this part, the purpose protein is NOP1v.This is a small peptide that can prevent the plants from accepting ethylene. In our experiment, we used it to inhibit the acceptance of ethylene and aging of cut flowers. We could not get NOP1 sufficiently when using BBa_K3815038, so we added a valine to the N-terminus of NOP1 according to N-end-rule.
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<h3><font size="4.5">Descriotion of this part</font> </h3>
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<h3><font size="3">Targeted protein</font> </h3>
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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 ''<partinfo>BBa_K3815009</partinfo>''. 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.
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<h3><font size="3">Purification system</font> </h3>
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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 Gyr 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.<br>
  
 
<!-- Add more about the biology of this part here
 
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Revision as of 12:33, 20 October 2021


NOP1v-Mxe GryA intein-PT-linker-His tag

Descriotion 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_K3815009. 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.

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 Gyr 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.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 120
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 120
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 120
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 120
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 120
    Illegal NgoMIV site found at 553
  • 1000
    COMPATIBLE WITH RFC[1000]