In constructing our Anaerobic Response System, we chose Invasin and Listeriolysin as part of our drug delivery system, building upon a construct from the Warsaw 2010 team, BBa_K299812. BBa_K299812 is a useful intermediate part containing the Invasin and Listeriolysin with ribosome binding sites in front of the genes. Different promoter sequences could be prepended ahead of the part and modified for our use.
However, when sequencing the part we received from the repository, our team found that the part sequence had major mismatches with the documented sequence. This was particularly serious for the invasin gene, which had only 55% identity between the translated amino acid sequences from the part wiki and our sequencing results. Furthermore, even though BLAST results showed that the invasin gene was indeed present, the strain of origin seemed to be different from the documentation. Lastly, the part does not strictly adhere to BioBricks standard, because the suffix sequence had only the PstI site left. (Further details and updated sequence can be found
here.)
As our lab only has the resources to functionally characterise the effect of the expression of invasin through an invasion assay, invasin was our main focus for the construction of parts for our Anaerobic Response System.
