Difference between revisions of "Part:BBa K320006"
 
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This BioBrick allows the expression of the Immunotoxin ( <partinfo>BBa_K320001</partinfo>) regulated by a strong constitutive promoter (<partinfo>BBa_K320002</partinfo>).
 
This BioBrick allows the expression of the Immunotoxin ( <partinfo>BBa_K320001</partinfo>) regulated by a strong constitutive promoter (<partinfo>BBa_K320002</partinfo>).
  
−
The Immunotoxin BioBricks (<partinfo>BBa_K320001</partinfo>) 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.
+
The promoter is constitutive promoter and the original BioBricks (<partinfo>BBa_K320002</partinfo>) also contain a ribosomal binding site for ''E. coli'' and ''Asaia''.  
  
−
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]
+
The Immunotoxin BioBrick (<partinfo>BBa_K320001</partinfo>) 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 ''P. falciparum''). This fragment is linked to the second part, the lytic peptide Shiva-1. Shiva-1 is a synthetic peptide, whose amino acid sequence is similar in length and amino acid properties to Cecropin B (38 amino acids), an antibacterial peptide. The purpose of the Immunotoxin is to specifically target and lyse the ookinete.
  
−
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.
+
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]
  
−
The BioBricks (<partinfo>BBa_K320001</partinfo>)contains a segments to export the immunotoxine protein to the periplasm.  
+
However ''E. coli'' is not a natural symbiont of the mosquito. We aim 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.
 +
 
 +
This BioBrick (<partinfo>BBa_K320001</partinfo>)also contains an export signal sequence to export the immunotoxine protein to the periplasm.  
  
  

Latest revision as of 18:25, 27 October 2010

Constitutive expression of Immunotoxin

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

The promoter is constitutive promoter and the original BioBricks (BBa_K320002) also contain a ribosomal binding site for E. coli and Asaia.

The Immunotoxin BioBrick (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 (sexual reproductive stage of a close relative of the Malaria parasite P. falciparum). This fragment is linked to the second part, the lytic peptide Shiva-1. Shiva-1 is a synthetic peptide, whose amino acid sequence is similar in length and amino acid properties to Cecropin B (38 amino acids), an antibacterial peptide. The purpose of the Immunotoxin is to specifically target and lyse the ookinete.

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 aim 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.

This BioBrick (BBa_K320001)also contains an export signal sequence 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]