Difference between revisions of "Part:BBa M36143:Experience"
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| + | 12/04/2015 | ||
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| + | 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 [https://parts.igem.org/Part:BBa_M36243] 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. | ||
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| + | [[File:ProRCells.jpg]][[File:MatureRCells.jpg]] | ||
Latest revision as of 06:04, 5 December 2015
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Applications of BBa_M36143
User Reviews
UNIQ1a72502c96ee9a01-partinfo-00000000-QINU UNIQ1a72502c96ee9a01-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.
