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

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.

SUBMITTER: Wierbowski SD 

PROVIDER: S-EPMC7260977 | biostudies-literature | 2020 May

REPOSITORIES: biostudies-literature

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

Wierbowski Shayne D SD   Vo Tommy V TV   Falter-Braun Pascal P   Jobe Timothy O TO   Kruse Lars H LH   Wei Xiaomu X   Liang Jin J   Meyer Michael J MJ   Akturk Nurten N   Rivera-Erick Christen A CA   Cordero Nicolas A NA   Paramo Mauricio I MI   Shayhidin Elnur E EE   Bertolotti Marta M   Tippens Nathaniel D ND   Akther Kazi K   Sharma Rita R   Katayose Yuichi Y   Salehi-Ashtiani Kourosh K   Hao Tong T   Ronald Pamela C PC   Ecker Joseph R JR   Schweitzer Peter A PA   Kikuchi Shoshi S   Mizuno Hiroshi H   Hill David E DE   Vidal Marc M   Moghe Gaurav D GD   McCouch Susan R SR   Yu Haiyuan H  

Proceedings of the National Academy of Sciences of the United States of America 20200512 21

Systematic mappings of protein interactome networks have provided invaluable functional information for numerous model organisms. Here we develop <u>P</u>CR-mediated <u>L</u>inkage of barcoded <u>A</u>dapters <u>T</u>o nucleic acid <u>E</u>lements for <u>seq</u>uencing (PLATE-seq) that serves as a general tool to rapidly sequence thousands of DNA elements. We validate its utility by generating the ORFeome for <i>Oryza sativa</i> covering 2,300 genes and constructing a high-quality protein-protei  ...[more]

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