A technology to quantitatively measure protein interactomes

2017 Fiscal Year Final Research Report

• Project/Area Number: 15K18466
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: System genome science
• Research Institution: The University of Tokyo
• Principal Investigator: Yachie Nozomu 東京大学, 先端科学技術研究センター, 准教授 (60636801)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Keywords: DNAバーコード / インタラクトーム / 超並列DNAシークエンシング / 酵母ツーハイブリッド / タンパク質間相互作用

Outline of Final Research Achievements

We remain one or two orders of magnitude away from a complete interaction map for humans and other major model organisms. Completion will require screening at substantially larger scales with many complementary assays, requiring further efficiency gains in proteome-scale interaction mapping. Here, we report Barcode Fusion Genetics-Yeast Two-Hybrid (BFG-Y2H), by which a full matrix of protein pairs can be screened in a single multiplexed strain pool. BFG-Y2H uses Cre recombination to fuse DNA barcodes from distinct plasmids, generating chimeric protein-pair barcodes that can be quantified via next-generation sequencing. We applied BFG-Y2H to four different matrices ranging in scale from ~25 K to 2.5 M protein pairs. The results show that BFG-Y2H increases the efficiency of protein matrix screening, with quality that is on par with state-of-the-art Y2H methods.

Free Research Field

合成生物学