Part:BBa_K320009
Constitutive expression of P28 in Asaia and Resistance to Tetracycline
This BioBrick makes bacteria resitant to Tetracycline, contains an origin of replication for Asaia/E. coli and drives constitutive expression of the protein P28.
The ookinete (Plasmodium stage in mosquito's gut) has been intensively studied by scientists, looking for an ideal transmission-blocking vaccine target. IGEM EPFL team's was interested in the ookinete surface membrane proteins Pfs25 and Pfs28 (link to other biobrick). It has been shown that the binding of specific antibodies to these surface proteins was fatal to more than 99% of the ookinetes. Indeed, P25 and P28 seem to be essential for the ookinete to be able to cross the midgut epithelium and thereafter transform into an oocyst.
Our team decided to design Asaia to express a soluble form of these 2 P-proteins. It could seem contra-intuitive to provide more of these proteins (by expression in Asaia), which are very important for the parasite survival, migration, and development. But our hypothesis is that adding a lot of these proteins in a soluble form will compete with the parasite surface protein and prevent the interaction necessary to its transmission. Because the functions of P25 and P28 are redundant, both proteins need to be inhibited or outcompeted to efficiently block the malaria transmission. More information can be found on our [http://2010.igem.org/wiki/index.php?title=Team:EPF_Lausanne/Project/Background website].
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 62
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 1934
Illegal AgeI site found at 2094 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 772
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