Part:BBa_K320001
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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 116
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 28
Illegal NgoMIV site found at 54
Illegal NgoMIV site found at 957 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 472
Illegal BsaI site found at 699
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