Part:BBa_M36143:Experience

This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

User Reviews

UNIQ0b897b0a9b687704-partinfo-00000000-QINU UNIQ0b897b0a9b687704-partinfo-00000001-QINU 12/04/2015

ProEP-B2 was used in comparison to MatureEP-B2 to test whether or not the ProEP-B2 inhibitor was necessary for the protein to be expressed. However, our fluorescence assay showed that MatureEP-B2 grew sufficiently in spite of not having an inhibitor, meaning that the inhibitor in ProEP-B2 was not necessary. A fluorescence assay was performed to test presence of ProEP-B2 in SDS lysed E.Coli. The fluorescence assay showed that ProEP-B2 was found to be in cells, since the reading detected increasing presence of 5-FAM quencher in the samples, a quencher that is cleaved by the ProEP-B2 protein. (See [1] for details on the ProEP-B2 composite results). A fluorescence assay of the MatureEP-B2 in SDS lysed E.Coli was also performed, an assay that also successfully showed the presence of MatureEP-B2 in cells. The MatureEP-B2 was expressed at much higher RLU than the ProEP-B2, so it is possible that the inhibitor was responsible for the smaller expression found in ProEP-B2 compared to MatureEP-B2 (graphs shown below for comparison). More testing is needed to conclude that the inhibitor was responsible for the difference in expression.