Coding

Part:BBa_K320001

Designed by: Perrudet Christian   Group: iGEM10_EPF_Lausanne   (2010-10-18)
Revision as of 15:49, 27 October 2010 by Gwen (Talk | contribs)

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Immunotoxin against P.Berghei

This BioBrick codes for an Immunotoxin. It is composed of two main parts. The first is a single-chain antibody fragment (scFv) directed to Pbs2l, a surface membrane protein of Plasmodium berghei ookinetes (sexual reproductive stage of a close relative of the Malaria parasite). This fragment is linked to the second part, a lytic peptide, Shiva-1. Shiva-1 is a synthetic peptide, which amino acids sequence of similar length (38 amino acids) and amino acid properties as Cecropin B, an antibacterial peptide. The purpose of the Immunotoxin is to specifically target and lyse the parasite.

Previous experiments showed that feeding mosquitoes with recombinant E. coli expressing this Immunotoxin induced a significantly lower oocyst density. [http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6T29-42JHDJD-9&_user=10&_coverDate=03%2F31%2F2001&_rdoc=1&_fmt=high&_orig=search&_origin=search&_sort=d&_docanchor=&view=c&_acct=C000050221&_version=1&_urlVersion=0&_userid=10&md5=5f4b78b08a1846e241faaed33bc76cb3&searchtype=a reference]

However E. coli is not a natural symbiont of the mosquito. We would like to improve this experiment by making Asaia express the immunotoxin in the mosquitoes intestinal tract, and run experiments to see if it significantly reduces the number of oocysts.

The BioBricks contains a segments to export the immunotoxin protein to the periplasm.

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 116
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 28
    Illegal NgoMIV site found at 54
    Illegal NgoMIV site found at 957
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 472
    Illegal BsaI site found at 699


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Categories
Parameters
None