Construction and activity assay of the activating transcription factor 3 reporter vector pATF/CRE-luc.
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.
Xu
Jun-Qing
JQ
Department of Biochemistry and Molecular Biology, Fourth Military Medical University, Xi'an 710032, China.
Deng
Jing-Lan
JL
Wu
You-Sheng
YS
Fu
Han-Yan
HY
Wang
Rui-Hua
RH
Zhang
Jian
J
Lu
Fan
F
Zhao
Zhong-Liang
ZL
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*
D051700:Activating Transcription Factor 3; D011994:Recombinant Proteins