Difference between revisions of "Part:BBa M36142:Experience"
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| + | 12/04/15 | ||
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| + | Mature EP-B2 was used in comparison to Pro EP-B2 to test whether the inhibitor sequence of the Pro EP-B2 was necessary for the protein to be made. Our fluorescence assay showed that mature EP-B2 fluoresced sufficiently meaning that the inhibitor sequence was not necessary. A fluorescence assay was performed to test presence of Mature EP-B2 in SDS lysed E.Coli. The fluorescence assay showed that Mature EP-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 Mature EP-B2 protein. Similar results were also found for the Mature EP-B2. The Mature EP-B2 actually 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:Pro EP-B2 Rhamnose Lysed.png]] | ||
| + | [[File:Mature EP-B2 Rhamnose Lysed.png]] | ||
Latest revision as of 07:11, 5 December 2015
This experience page is provided so that any user may enter their experience using this part.
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how you used this part and how it worked out.
Applications of BBa_M36142
User Reviews
UNIQ70fe02785e85caa8-partinfo-00000000-QINU UNIQ70fe02785e85caa8-partinfo-00000001-QINU 12/04/15
Mature EP-B2 was used in comparison to Pro EP-B2 to test whether the inhibitor sequence of the Pro EP-B2 was necessary for the protein to be made. Our fluorescence assay showed that mature EP-B2 fluoresced sufficiently meaning that the inhibitor sequence was not necessary. A fluorescence assay was performed to test presence of Mature EP-B2 in SDS lysed E.Coli. The fluorescence assay showed that Mature EP-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 Mature EP-B2 protein. Similar results were also found for the Mature EP-B2. The Mature EP-B2 actually 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.
