PYR1 Double-Hao and Triple Mutant Libraries
(Pooled Library #241288, #241289)

• Purpose:

  These two libraries are collections of designed PYR1 receptors in pBD, a Y2H vector, that can be used to isolate sensors using growth-based selections in yeast.

  PYR1-DSM-Hao is an improved version of the DSM (double site mutant) library from the Whitehead lab that purges a high number of constitutive mutants and reduces the number of wild-type receptors, enabling better coverage when screening and fewer constitutive (autoactivating) receptors. The PYR1-TSM is a triple-site mutation library as described in Tian (2025). It is constrained only to combine mutations known to influence ligand-receptor interactions and enables the isolation of sensors not accessible with the DSM-Hao or DSM libraries.

• Vector Backbone:

  pBD_PYR1_156 (Plasmid #241285)

• Depositing Labs: Sean Cutler
• Publication: Tian et al. bioRxiv 2025.05.15.654352. doi: 10.1101/2025.05.15.654352 .

Library Details

• Species: Arabidopsis
• Unique inserts:
  
  • DSM-Hao - 34,675
  • TSM - 324,352
• Size of insert: ~600 bp
• Expression: Yeast

Library Shipping

Each library is delivered in a microcentrifuge tube on blue ice. The tube's contents will not necessarily be frozen. For best results, minimize freeze/thaws.

• Volume: ∼15 µL
• Concentration: 100 ng/µL

Resource Information

• Protocols:
  • Library Amplification Protocol (DOCX, 18 KB) Please contact the depositing lab if additional protocol details are needed.
• Depositor Data:
  • List of mutations (TXT, 41.2 MB)

Terms and Licenses

Academic/Nonprofit Terms:

• UBMTA

Industry Terms:

• Not Available to Industry

Trademarks:

• Zeocin® is an InvivoGen trademark.

Depositor Comments

Figure 1: The coding sequence of PYR1 was divided into three blocks, each incorporating all designed mutations. Oligonucleotides containing these mutations were synthesized commercially, PCR-amplified, and assembled into a full-length PYR1 mutant library via Golden Gate cloning.

Please visit https://doi.org/10.1101/2025.05.15.654352 for bioRxiv preprint.