Protein Interaction Screening

Protein–protein interactions are crucial for forming protein complexes to carry out designated protein functions, such as the large machinery for DNA replication, the multi-protein-nucleic acid-complex of ribosome for translation, the nucleosome and associated epigenetic factors for gene expression regulation, and many more. The “interactome” is the core to any living cell, and the mapping of protein-protein interactions is the cornerstone to study their function. In addition, for drug development, knowing the molecular environment of a new drug target and its potential interaction partners in the cellular milieu can be predictive of functional roles and unwanted side effects. We offer a novel Next Generation Sequencing (NGS) Based Yeast Two Hybrid (Y2H) Protein Interaction Screening Service that overcomes false positives associated with traditional Y2H approaches and significant savings in time.

Key Features:

  • Higher throughput for better data coverage: NGS-Y2H procedure yielded at least a 10-fold more data coverage of prospective protein-protein interactions (PPIs) than traditional approach.
  • High quality data with low false positives: False positive generated by traditional Y2H can lead to unreliable data. Applying the NGS-Y2H procedure, we got at least a 10-fold better yield of prospective PPIs, while confounding false-positives can be excluded a priori.
  • Discover more novel interactions with advanced bioinformatics: Specialized bioinformatics algorithm for quick comparison across datasets and easy access to related experimental results.
  • Significant savings on time and cost: Higher quality screening data achieved upfront significantly simplifies downstream experimental design, shortens workflow, dramatically saves time to achieve more meaningful data and significantly lowers costs for downstream applications.

 

Applications:

  • A complete direct protein-protein interaction (binary interaction) map of a target protein of interest
  • Discover novel protein interaction partners under defined conditions
  • Amino acid variants/mutations affecting protein-protein interaction
  • Factors may affect protein-protein interaction

 

How to Order:

  • Customer supply: protein target to be studied
  • We deliver: Experimental design and execution, a comprehensive report and an optional web visualization of your data with advance bioinformatics tools
  • Quote today by fill the NGS-Y2H quotation form

Overview of Next Generation Protein Interaction Screening (NGIS)

Protein-protein interactions (PPIs) underlie virtually all biological processes and disease phenotypes, and hence are paramount for biomedical and biological research. Yeast two-hybrid (Y2H) is the most widely used technology for the exploration of PPIs and is established for high-throughput screening. In most Y2H experiments, complex prey cDNA libraries are screened with a known bait protein. Individual yeast clones that contain interacting bait-prey combinations are then enriched using auxotrophic markers over millions of non-interacting combinations.

In conventional Y2H, selected preys are then identified by Sanger sequencing. The one-by-one identification of single prey cDNAs limits the efficiency of the Y2H methodology, and is a serious cost constraint for the identification of large numbers of PPIs. Moreover, the clonal identification of Y2H interactions is by no means a quantitative measurement and does not allow for systematic comparisons between screens. Lack of quantitation and slow identification of PPIs via Y2H is a sharp contrast to the gathering of gene expression and protein-DNA interaction data by RNA-Seq and Chip-Seq.

We developed a screening and scoring scheme for the identification of Y2H PPIs from comprehensive yeast pools using next-generation sequencing (NGS). Replacing conventional screening and Sanger sequencing of individual clones with a pool-based selection and global identification has three major implications:

 

  • First, a massive increase in sequencing capacity that allows the identification of all positives from a Y2H screen at sharply reduced costs (up to 100,000-fold or more).
  • Second, the increased sequencing capacity allows screening at low stringency to address every potential interaction from the cDNA library, i.e. maximum sensitivity.
  • Third, NGS-Y2H allows the use of quantitative statistics to determine if a result is indeed bait-specific. The application of systematic controls allows a priori exclusion of false positive unspecific interactions, which used to be a major pitfall in conventional Y2H screens. Hence, quantitative scores are used for specificity.

 

Related Products:

•  Codon Optimization and Verification •  Gene-to-Protein Service
•  Gene-to-Antibody Service •  Protein Blotting/Staining System

 

 

Quotation & Ordering:

  • For quotation requests and questions, you may contact us by email, phone, or via our secured online message system.
  • To order, simply fill out the order form and submit to: order@protein-ct.com. Orders can be placed by phone, email, fax, or online with a formal PO (Purchase Order) . You may contact us anytime for assistance.

 

 
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