Difference between revisions of "Part:BBa K3802000"

(→‎Sequence and Features)
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This fusion protein aims to anchor MlrA, or microcystinase, an enzyme capable of degrading freshwater toxin microcystin-LR (MC-LR) to the outer membrane of  E. coli. This part  puts together a codon-optimized  subunit of  PgsA (poly-γ-glutamic acid synthetase) [https://parts.igem.org/Part:BBa_K2963020 (BBa_K2963020)], an anchoring motif to MlrA with 6x His [https://parts.igem.org/Part:BBa_K1907002 (BBa_K1907002)], with a 2 amino acid Gly-Ser linker [https://parts.igem.org/Part:BBa_J18920 (BBa_J18920)].  The addition of the His tag to MlrA gives this part the ability to be purified using Nickel affinity chromatography and detected using Western blotting with anti-His antibodies.  
 
This fusion protein aims to anchor MlrA, or microcystinase, an enzyme capable of degrading freshwater toxin microcystin-LR (MC-LR) to the outer membrane of  E. coli. This part  puts together a codon-optimized  subunit of  PgsA (poly-γ-glutamic acid synthetase) [https://parts.igem.org/Part:BBa_K2963020 (BBa_K2963020)], an anchoring motif to MlrA with 6x His [https://parts.igem.org/Part:BBa_K1907002 (BBa_K1907002)], with a 2 amino acid Gly-Ser linker [https://parts.igem.org/Part:BBa_J18920 (BBa_J18920)].  The addition of the His tag to MlrA gives this part the ability to be purified using Nickel affinity chromatography and detected using Western blotting with anti-His antibodies.  
 
This BioBrick was put under the control of an IPTG inducible ptac promoter [https://parts.igem.org/Part:BBa_K3802001 (BBa_K3802001)], as using a constitutive promoter could potentially lead to the depletion of resources for the bacteria.
 
This BioBrick was put under the control of an IPTG inducible ptac promoter [https://parts.igem.org/Part:BBa_K3802001 (BBa_K3802001)], as using a constitutive promoter could potentially lead to the depletion of resources for the bacteria.
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<partinfo>K3802000 SequenceAndFeatures</partinfo>

Revision as of 16:09, 21 October 2021

PgsA-MlrA fusion protein

Sequence and Features

This fusion protein aims to anchor MlrA, or microcystinase, an enzyme capable of degrading freshwater toxin microcystin-LR (MC-LR) to the outer membrane of E. coli. This part puts together a codon-optimized subunit of PgsA (poly-γ-glutamic acid synthetase) (BBa_K2963020), an anchoring motif to MlrA with 6x His (BBa_K1907002), with a 2 amino acid Gly-Ser linker (BBa_J18920). The addition of the His tag to MlrA gives this part the ability to be purified using Nickel affinity chromatography and detected using Western blotting with anti-His antibodies. This BioBrick was put under the control of an IPTG inducible ptac promoter (BBa_K3802001), as using a constitutive promoter could potentially lead to the depletion of resources for the bacteria.


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1792
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 827
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 813
    Illegal NgoMIV site found at 2323
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 304
    Illegal BsaI.rc site found at 2160