Dox-Inducible Expression of ddGFP PAR-T

The plasmids for Dox-inducible expression of the ddGFP PAR-T constructs were generated using a cDNA for ddGFP-A (Addgene, 40286) or ddGFP-B (Addgene, 40287). cDNAs for the PAR binding domains were amplified from previously published pET19b constructs (Gibson et al., 2017 (link)). The cDNAs were assembled and cloned first into pCDNA3 and then into pInducer20 or pET19b using Gibson assembly (NEB, E2621). The split luciferase constructs were synthesized as gene blocks (Integrated DNA Technologies), and then cloned into the pInducer20 vectors using Gibson assembly.

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Challa S., Ryu K.W., Whitaker A.L., Abshier J.C., Camacho C.V, & Kraus W.L. (2022). Development and characterization of new tools for detecting poly(ADP-ribose) in vitro and in vivo. eLife, 11, e72464.

Publication 2022

Gibson assembly gene blocks

Cdnas Gene Luciferase Plasmids Vectors

Corresponding Organization :

Other organizations : The University of Texas Southwestern Medical Center

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Variable analysis

independent variables

Dox-inducible expression of the ddGFP PAR-T constructs
Expression of the split luciferase constructs

dependent variables

Measured outcomes of the Dox-inducible expression of the ddGFP PAR-T constructs
Measured outcomes of the expression of the split luciferase constructs

control variables

CDNA for ddGFP-A (Addgene, 40286)
CDNA for ddGFP-B (Addgene, 40287)
PAR binding domains amplified from previously published pET19b constructs (Gibson et al., 2017)
Assembly and cloning of cDNAs into pCDNA3, pInducer20, and pET19b using Gibson assembly (NEB, E2621)

controls

Positive controls: Not explicitly mentioned
Negative controls: Not explicitly mentioned

Annotations

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