Difference between revisions of "Part:BBa K2365041"
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<partinfo>BBa_K2365041 short</partinfo> | <partinfo>BBa_K2365041 short</partinfo> | ||
| − | a Constitutive promoter from Saccharomyces cerevisiae | + | a Constitutive improved promoter from Saccharomyces cerevisiae |
Glyceraldehyde-3-phosphate dehydrogenase, -3- three glyceraldehyde dehydrogenase promoter (tdh1, tdh2, tdh3, tdh1) for weak promoter, tdh3 promoter, 3 phosphoglyceraldehyde catalytic oxidation (dehydrogenation) and phosphorylation of 1,3- - generation, is the central link of glucose metabolism, plays an important role in sugar metabolism. | Glyceraldehyde-3-phosphate dehydrogenase, -3- three glyceraldehyde dehydrogenase promoter (tdh1, tdh2, tdh3, tdh1) for weak promoter, tdh3 promoter, 3 phosphoglyceraldehyde catalytic oxidation (dehydrogenation) and phosphorylation of 1,3- - generation, is the central link of glucose metabolism, plays an important role in sugar metabolism. | ||
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| + | [[File:TDH3.jpg|600px|center]] | ||
| + | TDH3 from kit plate,the construct was transfected into Saccharomyces cerevisiae cells, the green fluorescence was observed under fluorescence microscope clearly. It indicates this part could work. | ||
| + | [[File:Yeast tutu.png|600px|center]] | ||
| + | Truncated TDH3 promoter,the construct was transfected into Saccharomyces cerevisiae cells, the green fluorescence was observed under fluorescence microscope clearly. | ||
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| − | Improved design: | + | ====Improved design:==== |
We construct the device： | We construct the device： | ||
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TDH3 promoter is a truncated promoter, GFP is a mutant. These is a invariable sequence upstream of GFP coding sequence. | TDH3 promoter is a truncated promoter, GFP is a mutant. These is a invariable sequence upstream of GFP coding sequence. | ||
| + | ====Characterization：==== | ||
| − | + | [[File:TDH3 IMPROVED.jpeg|450px|center]] | |
| + | Figure.The responses of promoter strengths of artificial TDH3 promoter and native promoter TDH3 were comprehensively compared. The strength of improved TDH3 was always higher than native promoter TDH3，but varied along with the genetic background of host. | ||
| − | + | [[File:启动子全部.jpeg|450px|center]] | |
| − | + | Figure.The fluorescence of the four test devices and control transformed into Saccharomyces cerevisiae and inoculated in YPD broth was measured after 8h. | |
| + | Nevertheless, the attractiveness of yeast as a platform organism (especially for metabolic engineering applications) needs continued efforts to expand the set of tools available. | ||
Latest revision as of 02:21, 2 November 2017
TDH3 promoter
a Constitutive improved promoter from Saccharomyces cerevisiae
Glyceraldehyde-3-phosphate dehydrogenase, -3- three glyceraldehyde dehydrogenase promoter (tdh1, tdh2, tdh3, tdh1) for weak promoter, tdh3 promoter, 3 phosphoglyceraldehyde catalytic oxidation (dehydrogenation) and phosphorylation of 1,3- - generation, is the central link of glucose metabolism, plays an important role in sugar metabolism.
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]
TDH3 from kit plate,the construct was transfected into Saccharomyces cerevisiae cells, the green fluorescence was observed under fluorescence microscope clearly. It indicates this part could work.
Truncated TDH3 promoter,the construct was transfected into Saccharomyces cerevisiae cells, the green fluorescence was observed under fluorescence microscope clearly.
Improved design:
We construct the device： TDH3 promoter+GFP+ADH1 terminator. TDH3 promoter is a truncated promoter, GFP is a mutant. These is a invariable sequence upstream of GFP coding sequence.
Characterization：
Figure.The responses of promoter strengths of artificial TDH3 promoter and native promoter TDH3 were comprehensively compared. The strength of improved TDH3 was always higher than native promoter TDH3，but varied along with the genetic background of host.
Figure.The fluorescence of the four test devices and control transformed into Saccharomyces cerevisiae and inoculated in YPD broth was measured after 8h. Nevertheless, the attractiveness of yeast as a platform organism (especially for metabolic engineering applications) needs continued efforts to expand the set of tools available.