Related Experiment Video
Updated: Aug 20, 2026
High-throughput Functional Screening using a Homemade Dual-glow Luciferase Assay
Published on: June 1, 2014
[Construction and identification of recombinant firefly luciferase reporter plasmid pGL2-PEPCK-Luc]
1Department of Endocrinology, PUMC Hospital, CAMS and PUMC, Beijing 100730, China.
Objective:
To evaluate in vitro regulation of phosphoenolpyruvate carboxykinase (PEPCK) gene promoter on gene transcription, and construct luciferase reporter plasmid pGL2-PEPCK-Luc.
Methods:
A 550 bp fragment of PEPCK promoter cut from plasmid pPEPCK-int was inserted into transitional vector PBS-SK to construct a transition plasmid PBS-PEPCK. Then the recombinant luciferase reporter plasmid pGL2-PEPCK-Luc was cloned.
Results:
Restriction enzymes and nucleotide sequence conformed that the coupling site of recombinant plasmid was correct without base mutation and deletion, and the sequence inserted was the same as data of GeneBank. The luciferase could be expressed in hepatoma cell transfected by pGL2-PEPCK-Luc.
Conclusion:
Established a new means to study transcriptional regulation of PEPCK promoter.
