promoterless control vector pgl3-basic  (Promega)

Journal: PLoS ONE

Article Title: Association of a Human FABP1 Gene Promoter Region Polymorphism with Altered Serum Triglyceride Levels

doi: 10.1371/journal.pone.0139417

Figure Lengend Snippet: Cells were co-transfected with 10 ng of the Renilla luciferase expression vector pRL-SV40 and 0.2 μg each of the pGL3-rs2919872G and pGL3-rs2919872A plasmids; the promoterless pGL3-Basic vector served as the negative control. Intracellular luciferase activity was measured 48 h after transfection. The relative luciferase units (RLU) were determined by comparison with the promoterless pGL3-Basic plasmid, which was assigned an arbitrary value of 1. Each transfection was performed in duplicate and the data are expressed as the mean ± SD of three separate experiments. (* P < 0.05).

Article Snippet: Cells were seeded (2×10 5 cells/well) in 12-well plates and transfected with the empty pGL3-Basic vector (a promoterless control) or with pGL3- rs2919872G or pGL3- rs2919872 A constructs using Lipofectamine 2000 (Invitrogen).

Techniques: Transfection, Luciferase, Expressing, Plasmid Preparation, Negative Control, Activity Assay

Journal:

Article Title: Gata4 Regulates Testis Expression of Dmrt1

doi: 10.1128/MCB.24.1.377-388.2004

Figure Lengend Snippet: Dmrt1 promoter activity is highest in primary cultures of Sertoli cells. Dmrt1(−5,000/+75)Luc was transfected into various cell types together with pRL-TK. The data are represented as the firefly/Renilla luciferase activity ratio of Dmrt1(−5,000/+75)Luc relative to the firefly/Renilla luciferase activity ratio of pGL3-Basic. Transfections were done a minimum of three times. Error bars represent the standard errors of the means. The inset shows an RNase protection analysis for Dmrt1 mRNA. RNA samples were isolated from primary rat Sertoli cells (SC), Sertoli cell lines MSC-1 and TM4, and primary myoid cells. tRNA was added as a negative control.

Article Snippet: Unless otherwise stated, myoid cells and JEG3 cells were transfected with 0.1 and 0.2 μg of promoterless control vector pGL3-Basic (Promega Corp., Madison, Wis.) or equimolar amounts of Dmrt1-luciferase constructs.

Techniques: Activity Assay, Transfection, Luciferase, Isolation, Negative Control

Journal:

Article Title: Gata4 Regulates Testis Expression of Dmrt1

doi: 10.1128/MCB.24.1.377-388.2004

Figure Lengend Snippet: Two regulatory regions are important for Dmrt1 transcription. (A) Various 5′-deletion mutants were generated from Dmrt1(−5,000/+75)Luc and characterized by transient-transfection analysis in either primary Sertoli cells or TM4 cells. The data are represented as the firefly/Renilla luciferase activity ratio of each construct relative to the firefly/Renilla luciferase activity ratio of pGL3-Basic (black bars are for primary Sertoli cells and white bars are for TM4 cells). Control represents the pGL3-Control vector, which contains the SV40 promoter and enhancer sequences. The −400/+75, −300/+75, −221/+75, −179/+75, and −75/+75 constructs were not tested in TM4 cells. Transfections were done a minimum of three times. Error bars represent the standard errors of the means. (B) Schematic of the Dmrt1 promoter region. Results in panel A identified distal (bp −3280/−2000) and proximal (below bp −150) regulatory regions that contribute to activity of the Dmrt1 promoter in primary Sertoli cells.

Article Snippet: Unless otherwise stated, myoid cells and JEG3 cells were transfected with 0.1 and 0.2 μg of promoterless control vector pGL3-Basic (Promega Corp., Madison, Wis.) or equimolar amounts of Dmrt1-luciferase constructs.

Techniques: Generated, Transfection, Luciferase, Activity Assay, Construct, Plasmid Preparation