Difference between revisions of "Part:BBa K3815005"
 
(12 intermediate revisions by the same user not shown)
Line 2: Line 2:
 
__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K3815005 short</partinfo>
 
<partinfo>BBa_K3815005 short</partinfo>
−
<h3><font size="4.5">Descriotion of this part</font> </h3>
+
==Description of this part==
 
<h3><font size="3">Targeted protein</font> </h3>
 
<h3><font size="3">Targeted protein</font> </h3>
−
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_K3815004</partinfo>'', so we added a valine to the N-terminus of NOP1 according to N-end-rule.
+
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_K3815004</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.
  
 
Sequence and Features<br><br>
 
Sequence and Features<br><br>
 
<h3><font size="3">Purification system</font> </h3>
 
<h3><font size="3">Purification system</font> </h3>
−
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>
+
[[File:ELK16.png|300px|thumb|right|Fig2.The mechanism of ELK16]]
 +
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.<br><br>
 +
We designed this part, however, we did not actually produce it with this part. We made this peptide with ''<partinfo>BBa_K3815010</partinfo>''.
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here
Line 22: Line 24:
 
<partinfo>BBa_K3815005 parameters</partinfo>
 
<partinfo>BBa_K3815005 parameters</partinfo>
 
<!-- -->
 
<!-- -->
 +
==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<br>
 +
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.
 +
<br>
 +
3.Tobias, J.W., Shrader, T.E., Rocap, G., and Varshavsky, A. (1991). The N-end rule in bacteria. Science 254, 1374–1377.<br>

Latest revision as of 18:59, 21 October 2021


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

Fig2.The mechanism of ELK16

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


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]


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.