Theranostics 2014; 4(8):808-822. doi:10.7150/thno.8255

Research Paper

Exploration of Panviral Proteome: High-Throughput Cloning and Functional Implications in Virus-host Interactions

Xiaobo Yu1*, Xiaofang Bian1*, Andrea Throop1, Lusheng Song1, Lerys Del Moral1, Jin Park1, Catherine Seiler1, Michael Fiacco1, Jason Steel1, Preston Hunter1, Justin Saul1, Jie Wang1, Ji Qiu1, James M. Pipas2, Joshua LaBaer1✉

1. The Virginia G. Piper Center for Personalized Diagnostics, Biodesign Institute, Arizona State University, Tempe, AZ 85287, USA
2. Department of Biological Sciences, University of Pittsburgh, Pennsylvania 15260, USA.
* These authors contributed equally to this work.

Abstract

Throughout the long history of virus-host co-evolution, viruses have developed delicate strategies to facilitate their invasion and replication of their genome, while silencing the host immune responses through various mechanisms. The systematic characterization of viral protein-host interactions would yield invaluable information in the understanding of viral invasion/evasion, diagnosis and therapeutic treatment of a viral infection, and mechanisms of host biology. With more than 2,000 viral genomes sequenced, only a small percent of them are well investigated. The access of these viral open reading frames (ORFs) in a flexible cloning format would greatly facilitate both in vitro and in vivo virus-host interaction studies. However, the overall progress of viral ORF cloning has been slow. To facilitate viral studies, we are releasing the initiation of our panviral proteome collection of 2,035 ORF clones from 830 viral genes in the Gateway® recombinational cloning system. Here, we demonstrate several uses of our viral collection including highly efficient production of viral proteins using human cell-free expression system in vitro, global identification of host targets for rubella virus using Nucleic Acid Programmable Protein Arrays (NAPPA) containing 10,000 unique human proteins, and detection of host serological responses using micro-fluidic multiplexed immunoassays. The studies presented here begin to elucidate host-viral protein interactions with our systemic utilization of viral ORFs, high-throughput cloning, and proteomic technologies. These valuable plasmid resources will be available to the research community to enable continued viral functional studies.

Keywords: Panviral proteome, ORFeome, Gateway® cloning, Gene synthesis, Virus-host interactions, Nucleic Acid Programmable Protein Arrays (NAPPA).

This is an open access article distributed under the terms of the Creative Commons Attribution (CC BY-NC) License. See http://ivyspring.com/terms for full terms and conditions.
How to cite this article:
Yu X, Bian X, Throop A, Song L, Moral LD, Park J, Seiler C, Fiacco M, Steel J, Hunter P, Saul J, Wang J, Qiu J, Pipas JM, LaBaer J. Exploration of Panviral Proteome: High-Throughput Cloning and Functional Implications in Virus-host Interactions. Theranostics 2014; 4(8):808-822. doi:10.7150/thno.8255. Available from http://www.thno.org/v04p0808.htm