Coding

Part:BBa_K2548009

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


SpALF4

Anti-lipopolysaccharide factors (ALFs) are antimicrobial peptides with a broad spectrum of antimicrobial activity. SpALF4 is an ALF homolog identified from Scylla paramamosain (Mud crab) that has shown antimicrobial activity against A. hydrophila.

Characterization

We examined the length of protein we extracted by performing SDS-PAGE to the supernatant after ultrasonic lysis. The protein size of SpALF4 is 11.54 kDa. Protein band in the correct molecular weight range were visualized in the area between 15 kDa and 10 kDa. There was no protein band in the same area for control group, indicating that SpALF4 was successfully 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 SpALF4


Figure 2 shows a group of broken A.hydrophila that has been processed by SpALF4. 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 SpALF4 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
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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