Difference between revisions of "Part:BBa K118025:Experience"
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The limonene generator with lac promoter (<partinfo>BBa_K118025</partinfo>) includes the limonene generator (<partinfo>BBa_K118024</partinfo>) under the control of a lac promoter (<partinfo>BBa_J33207</partinfo>). This composite part's purpose is to express the limonene synthase (<partinfo>BBa_I742110</partinfo>), which is also available with a ribosome site (<partinfo>BBa_I742111</partinfo>). | The limonene generator with lac promoter (<partinfo>BBa_K118025</partinfo>) includes the limonene generator (<partinfo>BBa_K118024</partinfo>) under the control of a lac promoter (<partinfo>BBa_J33207</partinfo>). This composite part's purpose is to express the limonene synthase (<partinfo>BBa_I742110</partinfo>), which is also available with a ribosome site (<partinfo>BBa_I742111</partinfo>). | ||
| − | [[Image:Ubcigem2011limonenegcms.jpg | frame | center | '''Gas Chromatography Mass Spectrum of (+)-Limonene:''' We expressed the limonene generator with lac promoter (<partinfo>BBa_K118025</partinfo>) in C41 DE3 ''E. coli'' and lysed the cell culture to obtain unpurified protein extract. The crude extract was used in an ''in vitro'' enzymatic assay to produce limonene from geranyl diphosphate (GPP). The samples were prepared and analyzed by gas chromatography mass spectrometry. | + | [[Image:Ubcigem2011limonenegcms.jpg | frame | center | '''Gas Chromatography Mass Spectrum of (+)-Limonene:''' We expressed the limonene generator with lac promoter (<partinfo>BBa_K118025</partinfo>) in C41 DE3 ''E. coli'' and lysed the cell culture to obtain unpurified protein extract. The crude extract was used in an ''in vitro'' enzymatic assay to produce limonene from geranyl diphosphate (GPP). The samples were prepared and analyzed by gas chromatography mass spectrometry. The gas chromatography retention time of 8.864 minutes (Figure A) and the mass spectrometry base peak at 93m/z (Figure B) are characteristic of limonene.]] |
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Revision as of 00:55, 14 September 2011
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Applications of BBa_K118025
Limonene synthase expression with pRARE2 in C41 DE3 E.coli that can be used to synthesize limonene terpenes.
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UNIQ421f7c316870ba5a-partinfo-00000000-QINU UNIQ421f7c316870ba5a-partinfo-00000001-QINU
Characterization
GC-MS analysis of Limonene Synthase function
The limonene generator with lac promoter (BBa_K118025) includes the limonene generator (BBa_K118024) under the control of a lac promoter (BBa_J33207). This composite part's purpose is to express the limonene synthase (BBa_I742110), which is also available with a ribosome site (BBa_I742111).
Gas Chromatography Mass Spectrum of (+)-Limonene: We expressed the limonene generator with lac promoter (BBa_K118025) in C41 DE3 E. coli and lysed the cell culture to obtain unpurified protein extract. The crude extract was used in an in vitro enzymatic assay to produce limonene from geranyl diphosphate (GPP). The samples were prepared and analyzed by gas chromatography mass spectrometry. The gas chromatography retention time of 8.864 minutes (Figure A) and the mass spectrometry base peak at 93m/z (Figure B) are characteristic of limonene.