Difference between revisions of "Part:BBa K1033112"
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It is believed that Uracil glucosyltransferase from Crocus Sativus (UGTCs2) cleaves products catalyzed by the enzyme zeaxanthin 7,8-cleavage dioxygenase (ZCD) and that it is neccesary for the production of crocin glucosides, safranal and picrocrocin. UGTCs2 was obtained from WashU 2012 in a synthetic construct togeher with ZCD. The genes were isolated by the use of primers with overhangs. The overhangs contained a RBS (B0034) and 6xHis-tag but overhangs without RBS were also used for isolation for future uses. We assembled with promoters but without success but when used mutagenesis we managed to attach several promoters of the J23-series. They were sequenced and tested in Western Blot, the sequences were correct but unfortunately we could not see a protein expression. This would show that we failed to make the Western Blot but the Sequencing shows that the promoter was correctly assembled. | It is believed that Uracil glucosyltransferase from Crocus Sativus (UGTCs2) cleaves products catalyzed by the enzyme zeaxanthin 7,8-cleavage dioxygenase (ZCD) and that it is neccesary for the production of crocin glucosides, safranal and picrocrocin. UGTCs2 was obtained from WashU 2012 in a synthetic construct togeher with ZCD. The genes were isolated by the use of primers with overhangs. The overhangs contained a RBS (B0034) and 6xHis-tag but overhangs without RBS were also used for isolation for future uses. We assembled with promoters but without success but when used mutagenesis we managed to attach several promoters of the J23-series. They were sequenced and tested in Western Blot, the sequences were correct but unfortunately we could not see a protein expression. This would show that we failed to make the Western Blot but the Sequencing shows that the promoter was correctly assembled. | ||
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| + | BBa_K2423002 is a modified version of this part. | ||
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Revision as of 13:06, 24 October 2017
Uracil glucosyltransferase (UGTCs2)
It is believed that Uracil glucosyltransferase from Crocus Sativus (UGTCs2) cleaves products catalyzed by the enzyme zeaxanthin 7,8-cleavage dioxygenase (ZCD) and that it is neccesary for the production of crocin glucosides, safranal and picrocrocin. UGTCs2 was obtained from WashU 2012 in a synthetic construct togeher with ZCD. The genes were isolated by the use of primers with overhangs. The overhangs contained a RBS (B0034) and 6xHis-tag but overhangs without RBS were also used for isolation for future uses. We assembled with promoters but without success but when used mutagenesis we managed to attach several promoters of the J23-series. They were sequenced and tested in Western Blot, the sequences were correct but unfortunately we could not see a protein expression. This would show that we failed to make the Western Blot but the Sequencing shows that the promoter was correctly assembled.
BBa_K2423002 is a modified version of this part.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]