Plant Yeast One-Hybrid Library Construction and Screening Service

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Plant Yeast One-Hybrid Library Construction and Screening Service

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CD BioSciences offers yeast one-hybrid (Y1H) technology for researching protein-DNA interactions. It combines transcriptional activation and detection systems in yeast cells for screening and characterizing plant proteins interacting with a given DNA sequence.

Y1H has shown great power in biological research. The application of the Y1H system has verified many known DNA-protein interactions while finding a variety of new transcription factors. Due to its sensitivity and reliability, this method has been widely used to clone specific transcription factors that are weak and difficult to purify by biochemical means.

The basic components of Y1H assay. Figure 1. The basic components of Y1H assay. (Reece-Hoyes, J. S., & Marian Walhout, A. J., 2012)

Service Content

Y1H library construction service

Steps Details Deliverables
RNA extraction RNA extraction, mRNA isolation.
  • Y1H Library glycerol bacteria 1.5mL (Titer > 1×107cfu/mL, fragment >1200bp, positive cloning rate > 95%).
  • Y1H library plasmid 1.5mL.
  • Electronic project report.
cDNA library construction
  • cDNA synthesis and junction ligation (three-frame library).
  • cDNA isolation homogenization by length (optional).
  • Recombination into yeast monohybrid vector pHis2/pAbAi and electro-transformation.
  • Library detection and plasmid extraction.
  • Data collation and analysis.

Y1H library screening service

Steps Details Deliverables
Promoter background screening Background screening of binding seq gene promoters.
  • Positive clone sequencing results.
  • Positive clone yeast plasmid.
  • Experimental result figures.
Library screening
  • Bait plasmid and library plasmid transformation.
  • Yeast screening and optimization of inhibitor concentration.
  • Sequencing of screening results.
Rotary verification
  • Screening results rotary validation
  • Data organization and analysis

Sample Requirements

Library construction

Plant cells > 1 x 107

Plant tissue > 2g

Total RNA > 200μg

Library screening

Bait gene sequence information or bait plasmids (pHis2-Bait or pAbAi-Bait).

Advantages of Y1H

  • Y1H can detect protein-DNA interactions that do not activate transcription. This includes proteins that do not directly play a role in transcription, such as replication proteins, DNA repair proteins, and blocking proteins.
  • Y1H is compatible with many existing libraries. Most Y1H experiments can use mixed prey libraries constructed for Y2H applications.
  • Y1H can detect isoform-specific interactions.

CD BioSciences has years of experience in library construction and screening, has completed thousands of various types of libraries, and can effectively guarantee the coverage of the library and library screening target gene detection. Please feel free to contact us to place your order.

References

  1. Cao, Y., et al. (2021). Construction of yeast one-hybrid library and screening of transcription factors regulating LhMYBSPLATTER expression in Asiatic hybrid lilies (Lilium spp.). BMC Plant Biology. 21(1), 563.
  2. Reece-Hoyes, J. S., & Marian Walhout, A. J. (2012). Yeast one-hybrid assays: a historical and technical perspective. Methods (San Diego, Calif.). 57(4), 441-447.

For research use only, not for clinical use.