title: Construction and activity assay of the activating transcription factor 3 reporter vector pATF/CRE-luc.
abstract: Activating transcription factor 3 (ATF3), a member of the activating transcription factor/cAMP responsive element binding protein (ATF/CREB) family of transcription factors, is induced by many physiological stresses. To investigate the activity of ATF/CREB in cells with physiological stresses, we developed a practical reporter vector, the plasmid pATF/CRE-luc, bearing activating transcription factor/cAMP responsive element (ATF/CRE) binding sites. This plasmid was constructed by inserting three repeats of the ATF/CRE binding element into the plasmid pG5luc, replacing the GAL-4 binding sites. The plasmids pACT/ATF3 and pATF/CRE-luc were transfected into HeLa and NIH3T3 cells, respectively, and the results showed that the expression of luciferase was increased in a dose-dependent manner on plasmid pACT/ATF3. The data suggested that the plasmid pATF/CRE-luc could be used as a sensitive and convenient reporter system of ATF3 activity.
authors.affiliation: Department of Biochemistry and Molecular Biology, Fourth Military Medical University, Xi'an 710032, China.
authors.forename: Jun-Qing
authors.initials: JQ
authors.lastname: Xu
authors.forename: Jing-Lan
authors.initials: JL
authors.lastname: Deng
authors.forename: You-Sheng
authors.initials: YS
authors.lastname: Wu
authors.forename: Han-Yan
authors.initials: HY
authors.lastname: Fu
authors.forename: Rui-Hua
authors.initials: RH
authors.lastname: Wang
authors.forename: Jian
authors.initials: J
authors.lastname: Zhang
authors.forename: Fan
authors.initials: F
authors.lastname: Lu
authors.forename: Zhong-Liang
authors.initials: ZL
authors.lastname: Zhao
mesh_terms: D051700:Activating Transcription Factor 3 / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D000818:Animals; D017930:Genes, Reporter / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D041681:NIH 3T3 Cells; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000378:metabolism; D014162:Transfection / Q000379:methods*
chemical_list: D051700:Activating Transcription Factor 3; D011994:Recombinant Proteins