Difference between revisions of "Part:BBa K2271118"
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==Characterization== | ==Characterization== | ||
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| − | We verified the expression of ValS via western blot. Therefore, we have a 3xFLAG-6xHis-Tag as a part in our plasmids, we can use for the antibodies. The estimated atomic mass of | + | We verified the expression of ValS via western blot. Therefore, we have a 3xFLAG-6xHis-Tag as a part in our plasmids, we can use for the antibodies. The estimated atomic mass of ValS is 69 kDa. |
| − | [[File:T--Cologne-Duesseldorf--western-blot-ValS.png|thumb|none | + | [[File:T--Cologne-Duesseldorf--western-blot-ValS.png|thumb|none| |
<strong>Protein abundance in WT and transformed cells from <i>Saccharomyces cerevisiae</i>:</strong> Protein abundance was detected using 6x His Tag Antibody. WT = wild type, ValS = Valencene Synthase, PTS1 = Peroxisom Targeting Signal 1]] | <strong>Protein abundance in WT and transformed cells from <i>Saccharomyces cerevisiae</i>:</strong> Protein abundance was detected using 6x His Tag Antibody. WT = wild type, ValS = Valencene Synthase, PTS1 = Peroxisom Targeting Signal 1]] | ||
Revision as of 18:33, 30 October 2017
ValS PTS1
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal suffix found in sequence at 2732
Illegal BglII site found at 860 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Usage and Biology
We used a valencene synthase(ValS) as part of our nootkatone pathway. ValS converts FPP into Valencene. We chose the ValS from Callitropis nootkatensis, because other valencene synthases perform relatively inefficient in microorganisms[1]. For example, they show a relatively poor product specificity, are less efficient in producing sesquiterpene than other synthases and have a significant amount of side products like germacrene A. In contrast to this, ValS from C. nootkatensis has a good product specificity and is more robust in terms of pH and temperature changes. It has also a better yield than citrus valencene synthases in yeast.
This part is designed for cytosolic expression, for peroxisomal import please choose part: BBa K2271124
Characterization
We verified the expression of ValS via western blot. Therefore, we have a 3xFLAG-6xHis-Tag as a part in our plasmids, we can use for the antibodies. The estimated atomic mass of ValS is 69 kDa.
