Coding

Part:BBa_K1727001

Designed by: Ying Kuan   Group: iGEM15_TCU_Taiwan   (2015-09-02)

Epinecidin-1 (Epinephelus coioides)


Mechanism of Epinecidin-1

Epinecidin-1 is a kind of antimicrobial peptide (AMPs) and it is a stable peptide that has extensive abilities to kill or inhibit the growth of bacteria. It plays a role in defense mechanism for Epinephelus coioides to against microbes. Epinecidin-1 use its chargeability to interact with bacteria cell membrane. Than use hydrophobic region interfere the membrane structure. This leads to cell lysis and bypasses bacterial antibiotic drug-resistance mechanisms. Moreover Epinecidin-1 has ability to help wound healing and has been proven by animal studies.

Features of Epinecidin-1

1. Small and simple: The sequence of Epinecidin-1 is only around 200 bp. For E.coli, 200 base pairs is in the working capacity. And its folding structure is a-helix it will not limited to E.coli translation system.

2. Activity: anti-Gram positive and negative bacteria, antiviral, anti-fungal, wound healing

3. Net charge: Its net charge is +3, so it can interact with bacterial cell membrane effectively.

4. Hydrophobic region: Epinecidin-1 hydrophobic region rate is 57 %, so it can mix with Vaseline into our medical dressing.


This part is a coding gene from the skin mucus of Epinephelus coioides.

TCU_TAIWAN_003_for_part.png

Fig.1 Gel electrophoresis for biobrick BBa_K1727001

Finally, we used EcoRI and PstI these two kinds of enzyme to check the insert really contain in the biobrick. We used 2% gel. The size of Epinecidin-1 insert should be 200 to 300 bp. There are bands on the sample 1~8 of enzyme digest product. As the result, we can make sure that the Epinecidin-1 has been ligated in backbone.


Disinfection ability of Epinecidin-1

2015tcutaiwanThe_peptides_efficiency_of_disinfection2.JPG
Fig.2 Chart of Epinecidin-1 disinfection ability to S. aureus

We had tested the disinfection efficiency of Epinecidin-1 on S. aureus to prove our system is work. As the chart we can see that Epinecidin-1 has ability to kill S. aureus. But the effect is not obvious. We speculated that the concentration is not enough to see the best efficiency. We will concentrate the peptides solution to do further experiments.


In vitro wound healing assay of HaCat cell line


Fig.1 HaCat cell tests for Epinecidin-1 Tcu_Taiwan_REsult_0919_3_1.jpeg
Fig.1-1 HaCat cell tests for Epinecidin-1 Tcu_Taiwan_REsult_0919_3_2.jpeg

In this group, Fig-1 shows there are no significant result, except Mock. The result of mock didn't show our expected. However, Fig. 1-1 has proven of significant effectiveness. When HaCat in 50ul of the Epinecidin-1 rarefied LB medium, the result indirect prove it has ability to help wound healing.

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 211
    Illegal BamHI site found at 1
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 154


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