A massively parallel barcoded sequencing pipeline enables generation of the first ORFeome and interactome map for rice

Proc Natl Acad Sci U S A. 2020 May 26;117(21):11836-11842. doi: 10.1073/pnas.1918068117. Epub 2020 May 12.

Abstract

Systematic mappings of protein interactome networks have provided invaluable functional information for numerous model organisms. Here we develop PCR-mediated Linkage of barcoded Adapters To nucleic acid Elements for sequencing (PLATE-seq) that serves as a general tool to rapidly sequence thousands of DNA elements. We validate its utility by generating the ORFeome for Oryza sativa covering 2,300 genes and constructing a high-quality protein-protein interactome map consisting of 322 interactions between 289 proteins, expanding the known interactions in rice by roughly 50%. Our work paves the way for high-throughput profiling of protein-protein interactions in a wide range of organisms.

Keywords: ORFeome; networks; next-generation sequencing; protein–protein interaction; rice.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Computational Biology / methods
  • DNA, Plant / genetics
  • Databases, Genetic
  • Genome, Plant / genetics
  • High-Throughput Nucleotide Sequencing / methods
  • Open Reading Frames / genetics*
  • Oryza / genetics*
  • Protein Interaction Mapping / methods*
  • Protein Interaction Maps / genetics*
  • Sequence Analysis, DNA / methods*

Substances

  • DNA, Plant