Difference between revisions of "Part:BBa K2686002"
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<partinfo>BBa_K2686002 parameters</partinfo>
 
<partinfo>BBa_K2686002 parameters</partinfo>
 
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==Results==
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The sequence was tested inside a a pet14 vector backbone inside an operon with a T7 promoter and a T7 terminator.
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===Self Assembly===
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To determine whether the protein cage is successfully expressed in a cell-free environment, multiple PAGE gels were run. The Encapsulin 60-mer has a mass of around 1.3 MDalton, which should reflect in a band very high up on the gels.
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===References===
 
===References===
 
Moon, H., Lee, J., Min, J. and Kang, S. (2014). Developing Genetically Engineered Encapsulin Protein Cage Nanoparticles as a Targeted Delivery Nanoplatform. Biomacromolecules, 15(10), pp.3794-3801.
 
Moon, H., Lee, J., Min, J. and Kang, S. (2014). Developing Genetically Engineered Encapsulin Protein Cage Nanoparticles as a Targeted Delivery Nanoplatform. Biomacromolecules, 15(10), pp.3794-3801.

Revision as of 13:49, 13 October 2018


Encapsulin protein with HexaHistidine insert

This is a BioBrick containing the sequence for Thermotoga maritima encapsulin, a bacterial protein nanocompartment which self assembles to form a 60-mer.

Usage and Biology

The part can be used to deliver cargo, both on the outer surface of the nanoparticle by fusing a peptide in between the 139/140 Amino Acids as well as the protein's C terminus. Cargo proteins can also be loaded inside the nano-cage using a tag binding to Encapsulin's interior surface. The protein is modified with an additonal amino acid sequence (GGGGGGHHHHHHGGGGG) between positions 43/44 granting it better stability and high heat resistance (Moon et al., 2014).

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 77
    Illegal BglII site found at 492
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 426
    Illegal SapI.rc site found at 457


Results

The sequence was tested inside a a pet14 vector backbone inside an operon with a T7 promoter and a T7 terminator.

Self Assembly

To determine whether the protein cage is successfully expressed in a cell-free environment, multiple PAGE gels were run. The Encapsulin 60-mer has a mass of around 1.3 MDalton, which should reflect in a band very high up on the gels.




References

Moon, H., Lee, J., Min, J. and Kang, S. (2014). Developing Genetically Engineered Encapsulin Protein Cage Nanoparticles as a Targeted Delivery Nanoplatform. Biomacromolecules, 15(10), pp.3794-3801.