Difference between revisions of "Part:BBa K3815001"
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==Purification==
 
==Purification==
 
[[File:Engineering 203 SDS-PAGE①.png|300px|thumb|right|SDS-PAGE of purified peptide]]
 
[[File:Engineering 203 SDS-PAGE①.png|300px|thumb|right|SDS-PAGE of purified peptide]]
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<h3><font size="4.5">Expression</font> </h3>
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<ul>
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<li>Cells were grown in 1000ml LB media at 27<sup>o</sup>.
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</ul>
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<ul>
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<li>Protein was expressed in 0.1mM IPTG for 2hours.
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</ul>
 
<h3><font size="4.5">Purification </font></h3>
 
<h3><font size="4.5">Purification </font></h3>
  

Revision as of 15:10, 19 October 2021


CecropinA-Mxe GryA intein-PT-linker-ELK16

Usage and Biology

This part is used for ELK16 system to purify the peptides. When this fused protein is produced, the part of ELK16 self-assembles and precipitates. After that, taking this aggregate and adding DTT to it, we cut the N terminal of Mxe GyrA intein. Then, we can get the targeted protein.In this part, the targeted protein is CecropinA derived from a moth. This is the antimicrobial peptide that can inhibit the growth of both gram-negative bacteria and gram-positive bacteria. In our experiment, we used it to kill the bacteria in the vase water.

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 814
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 67
    Illegal AgeI site found at 346
  • 1000
    COMPATIBLE WITH RFC[1000]

Purification

SDS-PAGE of purified peptide

Expression

  • Cells were grown in 1000ml LB media at 27o.
  • Protein was expressed in 0.1mM IPTG for 2hours.

Purification