The purpose of the Yeast One Hybrid Services Core is to carry out high-throughput mating-based yeast assays to screen for protein (transcription factor) – DNA (promoter or regulatory region) interactions. These gene-centered assays start with a DNA regulatory region of interest and enable identification of transcription factor(s) that interact with this region.
The Yeast One Hybrid Services Core provides a screening service between promoters and collections of Arabidopsis or Maize transcription factors.
Three collections of transcription factors are available:
i. A root stele-expressed collection of 800 Arabidopsis transcription factors (Gaudinier, Zhang et al. 2011; Li, Gaudinier et al. 2014) TFLIST 12-10-2014 CLM-1 (-850 root expressed TFs);
ii. A complete collection of 2,000 Arabidopsis transcription factors (Pruneda-Paz, Breton et al. 2014) TFLIST 11-30-2015 (full Arabidopsis TF collection);
iii. A collection of 500 maize transcription factors (Burdo, Gray et al. 2014) Maize TF Master list 3-2016.
The user will recombine their promoter(s) into a Gateway 5’TOPO plasmid (or Gateway pDONR-P4P1 plasmid if available) followed by the pMW2 and pMW3 yeast reporter plasmids. These constructs will then be provided to the UC Davis Yeast One Hybrid Services Core. The UC Davis Yeast One Hybrid Services Core completes yeast transformation, auto-activation tests, genotyping, and mating-based screens. Positive interactions are recorded for LacZ and HIS3 reporters.
The Yeast One Hybrid Service Core is located within the Brady Lab.
|Service||University of California||Non-Profit/ Education||For-Profit/ Industry|
|Y1H Screening of 1-14 Promoters/Arabidopsis 800||$6,129||$9,623||$11,817|
|Y1H Consumables only per Promoter/Arabidopsis 800||$138||$217||$266|
|Y1H Training Arabidopsis 800||$4,581||$7,192||$8,832|
|Y1H Addt’l Screening/Arabidopsis 800||$1,449||$2,275||$2,794|
|Y1H Screening of 1-6 Promoters/Arabidopsis 2000||$6,705||$10,527||$12,927|
|Y1H Consumables only per Promoter/Arabidopsis 2000||$374||$588||$722|
|Y1H Training Arabidopsis 2000||$5,157||$8,096||$9,942|
|Y1H Addt’l Screening/Arabidopsis 2000||$1,629||$2,557||$3,140|
|Y1H Screening of 1-24 Promoters/Maize 500||$6,561||$10,301||$12,650|
|Y1H Consumables only per Promoter/Maize 500||$104||$164||$201|
|Y1H Training Maize 500||$4,797||$7,531||$9,248|
|Y1H Addt’l Screening/Maize 500||$1,449||$2,275||$2,794|
|Labor Rate II||$36||$57||
Burdo, B., J. Gray, M. P. Goetting-Minesky, B. Wittler, M. Hunt, T. Li, D. Velliquette, J. Thomas, I. Gentzel, M. dos Santos Brito, M. K. Mejia-Guerra, L. N. Connolly, D. Qaisi, W. Li, M. I. Casas, A. I. Doseff and E. Grotewold (2014). “The Maize TFome–development of a transcription factor open reading frame collection for functional genomics.” Plant J 80(2): 356-366.
Gaudinier, A., L. Zhang, J. S. Reece-Hoyes, M. Taylor-Teeples, L. Pu, Z. Liu, G. Breton, J. L. Pruneda-Paz, D. Kim, S. A. Kay, A. J. Walhout, D. Ware and S. M. Brady (2011). “Enhanced Y1H assays for Arabidopsis.” Nat Methods 8(12): 1053-1055.
Li, B., A. Gaudinier, M. Tang, M. Taylor-Teeples, N. T. Nham, C. Ghaffari, D. Benson, M. Steinmann, J. A. Gray, S. M. Brady and D. J. Kliebenstein (2014). “Promoter based integration in plant defense regulation.” Plant Physiol.
Pruneda-Paz, J. L., G. Breton, D. H. Nagel, S. E. Kang, K. Bonaldi, C. J. Doherty, S. Ravelo, M. Galli, J. R. Ecker and S. A. Kay (2014). “A genome-scale resource for the functional characterization of Arabidopsis transcription factors.” Cell Rep 8(2): 622-632.