AMP. coli

        AMPs have an extensive ability in disinfect. Unlike antibiotics, AMPs use chargeability puncture the cell membrane to kill the bacteria therefore by passing bacterial antibiotic drug resistance mechanisms.[1] Two kinds of AMPs were selected as our reagents: Epinecidin-1 and Signiferin.
          Epinecidin-1 is a peptide comes from Epinephelus coioides, and Signiferin is comes from Crinia signifera. Both of them are extracted from the skin mucus. In addition, epinecidin-1 has the ability to help wounds healing and has been proven by animal studies.[2] Moreover, signiferin have great ability in disinfect Methicillin-Resistant Staphylococcus aureus (S. aureus), and had already been kindly proved by the TU-Delft 2013 iGEM team. [3]Combining these two properties, we believe that can alleviate the serious problem of skin injury.
        To produce AMPs and control AMPs expression, we apply the Lac operon and ligate the DNA of signal peptide into E. coli to help AMPs secret into culture medium. [4][5]Next, to prove that AMPs have the extensive ability in disinfection and helps the wound healing, selected cells and bacteria were tested in vitro, including the squamous epithelial cell and endothelial cell of the blood vessel and MRSA, and mice were used in vivo. Ultimately, create a wound dressing based on the above procedure.
       An excellent dressing made of AMPs will make a fast recovery.

Antimicrobial peptide

• Epinecidin-1:

1. From the skin mucus of Epinephelus coioides a kind of fish.

2. Has function of killing bacteria.

3. In addition, it has the ability to help wounds healing and has been proven by animal studies.

• Signiferin:

1. From the skin mucus of Crinia signifera a kind of tree frog.

2. Have function of killing bacteria.

3. Have great ability in disinfect Methicillin-Resistant Staphylococcus aureus (MRSA).

4. Had already been kindly proved by the 2013 TU-Delft iGEM team.

Signal peptide:

1. Helps AMPs to secret out of E. coli.

2. From Streptomyces lividans to trasport chitinase C to secretion system, which has been proven to work in E.coli

by reference.

Wound dressing:

Based on AMPs to develop into a potential material of wound dressing.

Reference

[1] Michael R. Yeaman & Nannette Y. Yount,..(2003) 'Mechanisms of Antimicrobial Peptide Action and Resistance' , Pharmacological Reviews,vol 55,p:27-55
[2] Han-Ning Huang, Chang-Jer Wu, Jyh-Yih Chen, Venugopal Rajanbabu , Chieh-Yu Pan, Yi-Lin Chan. Use of the antimicrobial peptide Epinecidin-1 to protect against MRSA infection in mice with skin injuries. Biomaterials 34 (2013) 10319e10327.
[3] V.M. Maselli, D. Bilusich, et al., Host-defence skin peptides of the Australian Streambank Froglet Crinia riparia: isolation and sequence determination by positive and negative ion electrospray mass spectrometry, Rapid Communications in Mass Spectrometry, Volume 20, Issue 5, pages 797–803, Mar 2006.
[4] Ken Tokuyasu, Satoshi Kaneko, Kiyoshi Hayashi, Yutaka Mori.Production of a recombinant chitin deacetylase in the culture medium of Escherichia coli cells.National Food Research Institute, Kannondai 2-1-2, Tsukuba 305-8642, Japan Received 26 July 1999
[5] TAKESHI FUJII, KIYOTAKA MIYASHITA. Multiple domain structure in a chitinase gene (chic) of Streptomyces lividans. Journal of General Microbiology (1993), 139, 677-686

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