Difference between revisions of "Part:BBa K517001"
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Results of detection by qRT-PCR were as follows | Results of detection by qRT-PCR were as follows | ||
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Medium Stock plasmid Average value SD | Medium Stock plasmid Average value SD | ||
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(qRT-PCR targeted to the loop portion of hairpin. Data was normalized with 25S rRNA. n=3 ) | (qRT-PCR targeted to the loop portion of hairpin. Data was normalized with 25S rRNA. n=3 ) | ||
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+ | [[File:プロモーター比較表.jpg|500px|thumb|center|'''Figure 1 : Results of detection by qRT-PCR (qRT-PCR targeted to the loop portion of hairpin. Data was normalized with 25S rRNA. n=3 )''']] | ||
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As a result, it was revealed that the constitutive GPD promoter had low RNA expression level. That level was even lower than from the conditional Gal1 promoter (BBa_J63006) in Glucose (Glu) medium, a condition which suppresses expression from Gal1promoter. We conducted another experiment with a longer 500 bp promoter which contains this GPD promoter sequence(<!--TDH3noyatu-->), and it was confirmed that the longer promoter has stronger expression. Therefore we cannot recommend using this part (BBa_K517001) for expressing protein excessively. | As a result, it was revealed that the constitutive GPD promoter had low RNA expression level. That level was even lower than from the conditional Gal1 promoter (BBa_J63006) in Glucose (Glu) medium, a condition which suppresses expression from Gal1promoter. We conducted another experiment with a longer 500 bp promoter which contains this GPD promoter sequence(<!--TDH3noyatu-->), and it was confirmed that the longer promoter has stronger expression. Therefore we cannot recommend using this part (BBa_K517001) for expressing protein excessively. |
Revision as of 09:16, 1 November 2017
GPD constitutive yeast promoter
Promoter for constitutively high expression.
Reference: This promoter is commonly used in several yeast vectors including the [http://www.addgene.org/yeast-gateway/ Advanced Gateway Vectors].
Book reference: Christine Guthrie, Gerald R. Fink. 2004. Guide to Yeast Genetics and Molecular and Cell Biology, Volume 1. Gulf Professional Publishing. Chapter 26: Vectors for Constitutive and Inducible Gene Expression in Yeast. Page 391.
Characterization by British Columbia iGEM 2011
The GPD promoter (BBa_K517001) and GAL promoter (BBa_K517000) as characterized by their regulation of the expression of a GFP reporter.
FACS Analysis of GFP expression as regulated by GPD and GAL Promoters
Fluorescence Analysis of GFP expression as regulated by GPD and GAL Promoters
Characterization by [http://2017.igem.org/Team:Kyoto iGEM Kyoto 2017]
We used this promoter in order to express long hairpin RNA in yeast. Long hairpin RNA targeting B. xylophilus AK1 mRNA or GFP mRNA was cloned downstream of the promoter and introduced into the 2-micron high copy number plasmid of budding yeast, and subsequently expressed in yeast. The Gal1 promoter ( BBa_J63006 ) was used as a control.
Results of detection by qRT-PCR were as follows
As a result, it was revealed that the constitutive GPD promoter had low RNA expression level. That level was even lower than from the conditional Gal1 promoter (BBa_J63006) in Glucose (Glu) medium, a condition which suppresses expression from Gal1promoter. We conducted another experiment with a longer 500 bp promoter which contains this GPD promoter sequence(), and it was confirmed that the longer promoter has stronger expression. Therefore we cannot recommend using this part (BBa_K517001) for expressing protein excessively.
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]