Plasmid Transfection Efficiency Optimization

The expression vector pcDNA3 was purchased from Invitrogen. An internal control plasmid for transfection efficiency, pRL-TK renilla luciferase (pRL-TK Luc), was purchased from Promega. The synthetic vitellogenin 3xERE-TATA fused to a luciferase reporter gene plasmid, 3xERE Luc, and a full-length human ERα expression plasmid, pcDNA/WT ERα, and pcDNA/SRC-2 expression plasmids have been described previously (51 (link)). All the Q375 mutant expression plasmids used were generated by Celplor Inc. (www.celplor.com) using the pENTR system.

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Li Y., Coons L.A., Houtman R., Carlson K.E., Martin T.A., Mayne C.G., Melchers D., Jefferson T.B., Ramsey J.T., Katzenellenbogen J.A, & Korach K.S. (2020). A mutant form of ERα associated with estrogen insensitivity affects the coupling between ligand binding and coactivator recruitment. Science signaling, 13(650), eaaw4653.

Publication 2020

pcDNA3 pRL-TK renilla luciferase (pRL-TK Luc),

Human Luciferase Plasmids Promega Renilla luciferase Reporter gene Transfection Vector Vitellogenin

Corresponding Organization :

Other organizations : National Institute of Environmental Health Sciences, National Institutes of Health, IBS Precision Engineering (Netherlands), University of Illinois Urbana-Champaign

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Variable analysis

independent variables

PcDNA3 expression vector
PRL-TK renilla luciferase (pRL-TK Luc) plasmid
3xERE Luc synthetic vitellogenin 3xERE-TATA fused to a luciferase reporter gene plasmid
PcDNA/WT ERα full-length human ERα expression plasmid
PcDNA/SRC-2 expression plasmid
Q375 mutant expression plasmids

dependent variables

Transfection efficiency
Luciferase reporter gene expression

control variables

Experimental conditions and procedures

positive controls

PRL-TK renilla luciferase (pRL-TK Luc) plasmid as an internal control for transfection efficiency

negative controls

Not explicitly mentioned

Annotations

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