Difference between revisions of "Part:BBa K517002:Experience"
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===Applications of BBa_K517002=== | ===Applications of BBa_K517002=== | ||
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| + | Alpha-pinene synthase expression with pRARE2 in C41 DE3 E.coli that can be used to synthesize alpha-pinene. | ||
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===User Reviews=== | ===User Reviews=== | ||
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| + | ==Characterization by British Columbia iGEM 2011== | ||
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| + | ===GC-MS analysis of Limonene Synthase function by British Columbia iGEM 2011=== | ||
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| + | [[Image:Ubcigem2011limonenegcms.jpg | frame | center | '''Gas Chromatography Mass Spectrum of Alpha-pinene Synthase Products:''' We expressed the alpha-pinene synthase (<partinfo>BBa_K517002</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 alpha-pinene from geranyl diphosphate (GPP) substrate. The samples were prepared and analyzed by gas chromatography mass spectrometry. (A) We analysed the products of the alpha-pinene synthase and found alpha-pinene, beta-pinene and small quantities of other monoterpenes. (B) Cross referencing with standards and a compound library revealed that each monoterpene product had the characteristic base peaks and retention time. The left panel shows the identification of alpha-pinene and the right panel shows the identification of beta-pinene as products of our alpha-pinene synthase.]] | ||
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Revision as of 21:02, 23 September 2011
This experience page is provided so that any user may enter their experience using this part.
Please enter
how you used this part and how it worked out.
Applications of BBa_K517002
Alpha-pinene synthase expression with pRARE2 in C41 DE3 E.coli that can be used to synthesize alpha-pinene.
User Reviews
UNIQ2e5a2ce998ccfd5f-partinfo-00000000-QINU UNIQ2e5a2ce998ccfd5f-partinfo-00000001-QINU
Characterization by British Columbia iGEM 2011
GC-MS analysis of Limonene Synthase function by British Columbia iGEM 2011
Gas Chromatography Mass Spectrum of Alpha-pinene Synthase Products: We expressed the alpha-pinene synthase (BBa_K517002) 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 alpha-pinene from geranyl diphosphate (GPP) substrate. The samples were prepared and analyzed by gas chromatography mass spectrometry. (A) We analysed the products of the alpha-pinene synthase and found alpha-pinene, beta-pinene and small quantities of other monoterpenes. (B) Cross referencing with standards and a compound library revealed that each monoterpene product had the characteristic base peaks and retention time. The left panel shows the identification of alpha-pinene and the right panel shows the identification of beta-pinene as products of our alpha-pinene synthase.