Part:BBa_K320006

Constitutive expression of Immunotoxin

This BioBrick allows the expression of the Immunotoxin ( BBa_K320001) regulated by a strong constitutive promoter (BBa_K320002).

The Immunotoxin BioBricks (BBa_K320001) 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 (xxx stadium 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 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 reference]

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 BioBricks (BBa_K320001)contains a segments to export the immunotoxine protein to the periplasm.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 62
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 883
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]