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PEXP is an endogenous promoter in Yarrowia lipolytica, which was first registered in 2020. In order to characterize the activity and strength of promoter PEXP, we use the Nluc analysis by constructing the recombinant plasmid pYXLP’-PEXP-Nluc. The constructed recombinant plasmid pYXLP’-PEXP-Nluc was further transformed into Y. lipolytica po1f. Next, we performed the shaking flask to test the Nanoluc luciferase analysis. The experimental result showed a clear fluorescent signal (Fig. 3), demonstrating that the promoter PEXP was able to drive gene transcription in Y. lipolytica and it can be considered as a moderately strong promoter. These results provide references for future iGEM team to select promoters. | PEXP is an endogenous promoter in Yarrowia lipolytica, which was first registered in 2020. In order to characterize the activity and strength of promoter PEXP, we use the Nluc analysis by constructing the recombinant plasmid pYXLP’-PEXP-Nluc. The constructed recombinant plasmid pYXLP’-PEXP-Nluc was further transformed into Y. lipolytica po1f. Next, we performed the shaking flask to test the Nanoluc luciferase analysis. The experimental result showed a clear fluorescent signal (Fig. 3), demonstrating that the promoter PEXP was able to drive gene transcription in Y. lipolytica and it can be considered as a moderately strong promoter. These results provide references for future iGEM team to select promoters. | ||
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Revision as of 15:02, 17 October 2021
EXP Promoter
EXP is a strong promoter from Yarrowia lipolytica. It can express genes efficiently. The EXP part will be cloned to a vector according to the iGEM standard design principle to express the glucoamylase or alpha-amylase genes.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal SpeI site found at 756
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 817
Illegal SpeI site found at 756 - 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal SpeI site found at 756
- 25INCOMPATIBLE WITH RFC[25]Illegal SpeI site found at 756
- 1000COMPATIBLE WITH RFC[1000]
Contribution From NNU-China 2021
Group: iGEM Team NNU-China 2021
Author: Yan Xu
Summary: Testing its transcription level
Characterization from iGEM21-NNU-China
PEXP is an endogenous promoter in Yarrowia lipolytica, which was first registered in 2020. In order to characterize the activity and strength of promoter PEXP, we use the Nluc analysis by constructing the recombinant plasmid pYXLP’-PEXP-Nluc. The constructed recombinant plasmid pYXLP’-PEXP-Nluc was further transformed into Y. lipolytica po1f. Next, we performed the shaking flask to test the Nanoluc luciferase analysis. The experimental result showed a clear fluorescent signal (Fig. 3), demonstrating that the promoter PEXP was able to drive gene transcription in Y. lipolytica and it can be considered as a moderately strong promoter. These results provide references for future iGEM team to select promoters.
- Fig.1 A. The cell growth of strain po1f-PEXP-Nluc P. B. Nanoluc luciferase was used to quantify the strength of promoter PEXP.