Coding

Part:BBa_K2548010

Designed by: Zhuoyi Yang   Group: iGEM18_BNDS_CHINA   (2018-10-01)


Scygonadin (codon optimized)

Scygonadin is an anionic antimicrobial peptide identified from the seminal plasma of the species Scylla serrata (Giant mud crab). It has shown the antimicrobial activity against A. hydrophila. Its DNA reveals that it is only expressed in male crabs, helping them to fight off bacterial invasions.

Characterization

We examined the length of protein we extracted by performing SDS-PAGE to the supernatant after ultrasonic lysis. The protein size of SCY is 12.54 kDa. Protein band in the correct molecular weight range was visualized in the area between 15 kDa and 10 kDa. There was no protein band in the same area for control group, indicating that SCY was expressed.

Fig.1 Protein bands of antimicrobial peptides

We tested the effect of different antimicrobial peptides through the procedures described in Experiments. The photo below show Aeromonas hydrophila under electron microscope.

Fig.2 Cells of A.hydrophila processed by SCY


Figure 2 shows a group of broken A.hydrophila that has been processed by SCY. The edges of the cells are clearly rougher than those of cells in the control group, with some obvious light leaves as circled in blue. We suppose the darkened parts among and besides the cells (circled in red) are the cell structures flowed out of the cell via the gaps on cell walls.

Through the electron microscope photos, we could demonstrate that our SCY solution caused damage on cell walls of A.hydrophila.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 73
  • 1000
    COMPATIBLE WITH RFC[1000]


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