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ARS Home » Northeast Area » Ithaca, New York » Robert W. Holley Center for Agriculture & Health » Plant, Soil and Nutrition Research » Research » Publications at this Location » Publication #284614
Title: Enhanced Y1H Assays for Arabidopis

Author
item GAUDINIER, ALLISON - University Of California
item ZHANG, LIFANG - Cold Spring Harbor Laboratory
item REECE-HOYES, JOHN - University Of Massachusetts
item TAYLOR-TEEPLES, MALLORIE - University Of California
item PU, LI - University Of California
item LIU, ZHIJIE - Cold Spring Harbor Laboratory
item BRETON, GHISLAIN - University Of California
item PRUNEDA-PAZ, JOSE - University Of California
item KIM, DAHAE - University Of California
item KAY, STEVE - University Of California
item WALHOUT, ALBERTHA - University Of Massachusetts
item Ware, Doreen
item BRADY, SIOBHAM - University Of California

Submitted to: Nature Methods
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 8/30/2011
Publication Date: 10/30/2011
Citation: Gaudinier, A., Zhang, L., Reece-Hoyes, J.S., Taylor-Teeples, M., Pu, L., Liu, Z., Breton, G., Pruneda-Paz, J.L., Kim, D., Kay, S.A., Walhout, A.J., Ware, D., Brady, S.M. 2011. Enhanced Y1H Assays for Arabidopis. Nature Methods. 8:1053-1055.

Interpretive Summary: An Arabidopsis full-length enhanced yeast one hybrid (eY1H) transcription factor (TF) resource comprised of 645 or 91% of TFs expressed in the root stele and 75% of root-expressed TFs is presented. We demonstrate that this resource and the eY1H assay enable rapid, efficient and systematic mapping of plant TF-promoter interactions using thirteen stele-expressed TF promoters. 158 interactions were identified, many of which occur physically or are regulatory in planta.

Technical Abstract: Transcription regulation plays a key role in development and response to environment. To understand this mechanism, we need to know which transcription factor (TFs) would bind to which promoter, thus regulate their target gene expression. Yeast one-hybrid (Y1H) technique can be used to map this kind of interaction. This paper described the improvement of this method as enhanced Y1H (eY1H), using robotic mating method to replace laborious transformation method, thus more efficient and reduce the cost. Most importantly, this paper provided a more complete TF resource (including 69 de novo cloning of TFs) that represent 91% of TFs that expressed in root stele and 73% of TFs expressed in root. It would be great resource to the plant community.