Difference between revisions of "Part:BBa K320001"

Line 2: Line 2:
 
<partinfo>BBa_K320001 short</partinfo>
 
<partinfo>BBa_K320001 short</partinfo>
  
This part contain a toxine (shiva) combined to an anticorps against Pbs21, a membrane protein of the plasmudium.
+
This Biobrick is 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 P. berghei ookinetes. This fragment is linked to the second part, a lytic peptide, Shiva-1. Shiva-1 is a synthetic peptide which amino acids sequence is similar in terms of length (38 amino acids) and amino acid properties to 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 Shigeto Yoshida, Bacteria expressing single-chain immunotoxin inhibit malaria parasite development in mosquitoes, Molecular and Biochemical Parasitology (2001)]
 +
 
 +
However E. Coli is not a natural symbiote of the mosquito. We would like to improve this experiment by making Asaia express the immunotoxin in the mosquitos intestinal tract, and run experiments to see if it significantly reduces the number of oocysts.
 +
 
 +
The sequence of the immunotoxin was taken from our reference paper [1], and the codons were optimized for E.Coli before ordering the sequence.  
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 16:48, 24 October 2010

Immunotoxin against P.Berghei

This Biobrick is 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 P. berghei ookinetes. This fragment is linked to the second part, a lytic peptide, Shiva-1. Shiva-1 is a synthetic peptide which amino acids sequence is similar in terms of length (38 amino acids) and amino acid properties to 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 Shigeto Yoshida, Bacteria expressing single-chain immunotoxin inhibit malaria parasite development in mosquitoes, Molecular and Biochemical Parasitology (2001)]

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

The sequence of the immunotoxin was taken from our reference paper [1], and the codons were optimized for E.Coli before ordering the sequence.

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