Difference between revisions of "Part:BBa K678004"

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Summary：in this part, we made contributions to the quantitative characterization data of mammalian PGK Promoter. We measured the strength of this promoter by the expression of firefly luciferase at different time.  
 
Summary：in this part, we made contributions to the quantitative characterization data of mammalian PGK Promoter. We measured the strength of this promoter by the expression of firefly luciferase at different time.  
  
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Characterization: To characterize the PGK promoter, firefly luciferase was used as the reporter gene to quantify the transcriptional strength of PGK promoter. To be specific, pmirGLO carrying PGK promoter was transfected into liver carcinoma cell line HepG2. 0, 6, 12 and 24h after transfection, cells were harvested for measurement of luciferase activity. Firefly luciferase activity was measured by Beyotime™  Dual Luciferase Reporter Gene Assay Kit. Results are shown below.  
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Characterization: To characterize the PGK promoter, firefly luciferase was used as the reporter gene to quantify the transcriptional strength of promoter. To be specific, pmirGLO carrying PGK promoter was transfected into liver carcinoma cell line HepG2. 0, 6, 12 and 24h after transfection, cells were harvested, and firefly luciferase activity was measured by Beyotime™  Dual Luciferase Reporter Gene Assay Kit. Results are shown below.  
  
 
https://static.igem.org/mediawiki/parts/b/b3/T--NUDT_CHINA--PGK2.jpg
 
https://static.igem.org/mediawiki/parts/b/b3/T--NUDT_CHINA--PGK2.jpg

Revision as of 18:18, 21 October 2019


PGK, mammalian promoter

Promoter for expression of genes in mammalian cells

Contribution: NUDT_CHINA 2019

Summary：in this part, we made contributions to the quantitative characterization data of mammalian PGK Promoter. We measured the strength of this promoter by the expression of firefly luciferase at different time.

Characterization: To characterize the PGK promoter, firefly luciferase was used as the reporter gene to quantify the transcriptional strength of promoter. To be specific, pmirGLO carrying PGK promoter was transfected into liver carcinoma cell line HepG2. 0, 6, 12 and 24h after transfection, cells were harvested, and firefly luciferase activity was measured by Beyotime™ Dual Luciferase Reporter Gene Assay Kit. Results are shown below.

T--NUDT_CHINA--PGK2.jpg


Figure 1. PGK-drived firefly luciferase activity.

The result showed that within 24h, transcription of the mammalian PGK promoter-drived luciferase level keeps rising and reached maximum at 24h after transfection.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal SpeI site found at 143
    Illegal PstI site found at 400
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal SpeI site found at 143
    Illegal PstI site found at 400
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal SpeI site found at 143
    Illegal PstI site found at 400
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal SpeI site found at 143
    Illegal PstI site found at 400
  • 1000
    COMPATIBLE WITH RFC[1000]