Constructing luciferase reporter and expression plasmids

The luciferase reporter harboring the TDAG51 promoter fragments containing nt –2,450 to +551 (P_T51-3K-Luc) was described previously (Park et al., 2013 (link)). For the construction of aP2 luciferase reporter (aP2-Luc), the aP2 promoter fragments from nt –5,379 to +21 were amplified by polymerase chain reaction (PCR) from the murine genomic DNA and subcloned into the pGL3-basic vector (Promega, USA). The eukaryotic expression plasmid of Flag-tagged TDAG51 (Flag-TDAG51) was described previously (Park et al., 2013 (link)). Flag-tagged RXRα (Flag-RXRα) was kindly provided by Yong-Ho Ahn (Yonsei University College of Medicine, Korea) (Kim et al., 2009 (link)). The epitope-tagged (Flag-, Myc-, Xp-, or GST-tagged) eukaryotic expression plasmids for TDAG51, PPARγ, RXRα, and their corresponding deletion mutants were generated by PCR amplification and subcloned into pFLAG-CMV2 (Sigma-Aldrich), pcDNA3.1/myc-His/lacZ (Invitrogen), pcDNA3.1-His (Invitrogen), pEBG (Park et al., 2015 (link)), and pMXs-puro (Park et al., 2015 (link)).

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Kim S., Lee N., Park E.S., Yun H., Ha T.U., Jeon H., Yu J., Choi S., Shin B., Yu J., Rhee S.D., Choi Y, & Rho J. (2020). T-Cell Death-Associated Gene 51 Is a Novel Negative Regulator of PPARγ That Inhibits PPARγ-RXRα Heterodimer Formation in Adipogenesis. Molecules and Cells, 44(1), 1-12.

Publication 2020

pGL3-basic vector pFLAG-CMV2 pcDNA3.1/myc-His/lacZ pcDNA3.1-His

Deletion Epitope Eukaryotic Genomic Lacz Luciferase Medicine Murine Pgl3 Plasmids Polymerase chain reaction Pparγ Promega Vector

Corresponding Organization :

Other organizations : Chungnam National University, Korea Research Institute of Chemical Technology, University of Pennsylvania

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Variable analysis

independent variables

Promoter fragments containing nt -2,450 to +551 of the TDAG51 promoter (P_T51-3K-Luc)
Promoter fragments from nt -5,379 to +21 of the aP2 promoter (aP2-Luc)
Flag-tagged TDAG51 (Flag-TDAG51)
Flag-tagged RXRα (Flag-RXRα)
Epitope-tagged (Flag-, Myc-, Xp-, or GST-tagged) eukaryotic expression plasmids for TDAG51, PPARγ, RXRα, and their corresponding deletion mutants

dependent variables

Luciferase reporter activity

control variables

PGL3-basic vector (Promega, USA)
PcDNA3.1/myc-His/lacZ (Invitrogen)
PcDNA3.1-His (Invitrogen)
PMXs-puro

controls

Positive control: None specified
Negative control: None specified

Annotations

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