Coding

Part:BBa_K3515002

Designed by: Kody Klupt, Sono Khan   Group: iGEM20_Queens_Canada   (2020-05-22)
Revision as of 14:25, 25 May 2020 by Kody klupt (Talk | contribs)

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Synechococcus Phosphate Binding Protein with cysteine modification(s) to bind to a biosensor.

Synechococcus Phosphate Binding Protein (PiBP) selectively binds inorganic phosphate in its active site, inducing a conformation change in the N and C termini regions, respectively. This makes it a distinguishable candidate for in vivo or in vitro phosphate monitoring using fluorescence resonance energy transfer (FRET). Coupling this protein with two fluorophores can permit phosphate detection. Phosphate detection is vital as phosphate is used in cells for the synthesis of DNA, phospholipids, ATP, and is involved in other signal cascades or post-transcriptional modifications. Phosphate is also a crucial biomarker used in clinical medicine for tracking the progression and status of Chronic Kidney Disease (CKD) patients. As such a biosensor for phosphate tracking may be of great interest to patients and clinicians. This part includes a mutated PiBP to have a cysteine modification that will bind cysteine linker arms and be used for biosensor immobilization allowing the detection of inorganic phosphate.

File3.png

Cysteine immobilization modifications of binding protein. Protein structures were obtained from the RCSB Protein Data Bank. All protein residues are shown in a blue cartoon preset with cysteine residues labelled in red cartoon, respectively, using PyMOL software.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 828
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 418
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
//chassis/prokaryote/ecoli
Parameters
biologySynechococcus