title: Exploration of panviral proteome: high-throughput cloning and functional implications in virus-host interactions.
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.
authors: 1. The Virginia G. Piper Center for Personalized Diagnostics, Biodesign Institute, Arizona State University, Tempe, AZ 85287, USA.
authors: Xiaobo
authors: X
authors: Yu
authors: 1. The Virginia G. Piper Center for Personalized Diagnostics, Biodesign Institute, Arizona State University, Tempe, AZ 85287, USA.
authors: Xiaofang
authors: X
authors: Bian
authors: 1. The Virginia G. Piper Center for Personalized Diagnostics, Biodesign Institute, Arizona State University, Tempe, AZ 85287, USA.
authors: Andrea
authors: A
authors: Throop
authors: 1. The Virginia G. Piper Center for Personalized Diagnostics, Biodesign Institute, Arizona State University, Tempe, AZ 85287, USA.
authors: Lusheng
authors: L
authors: Song
authors: 1. The Virginia G. Piper Center for Personalized Diagnostics, Biodesign Institute, Arizona State University, Tempe, AZ 85287, USA.
authors: Lerys Del
authors: LD
authors: Moral
authors: 1. The Virginia G. Piper Center for Personalized Diagnostics, Biodesign Institute, Arizona State University, Tempe, AZ 85287, USA.
authors: Jin
authors: J
authors: Park
authors: 1. The Virginia G. Piper Center for Personalized Diagnostics, Biodesign Institute, Arizona State University, Tempe, AZ 85287, USA.
authors: Catherine
authors: C
authors: Seiler
authors: 1. The Virginia G. Piper Center for Personalized Diagnostics, Biodesign Institute, Arizona State University, Tempe, AZ 85287, USA.
authors: Michael
authors: M
authors: Fiacco
authors: 1. The Virginia G. Piper Center for Personalized Diagnostics, Biodesign Institute, Arizona State University, Tempe, AZ 85287, USA.
authors: Jason
authors: J
authors: Steel
authors: 1. The Virginia G. Piper Center for Personalized Diagnostics, Biodesign Institute, Arizona State University, Tempe, AZ 85287, USA.
authors: Preston
authors: P
authors: Hunter
authors: 1. The Virginia G. Piper Center for Personalized Diagnostics, Biodesign Institute, Arizona State University, Tempe, AZ 85287, USA.
authors: Justin
authors: J
authors: Saul
authors: 1. The Virginia G. Piper Center for Personalized Diagnostics, Biodesign Institute, Arizona State University, Tempe, AZ 85287, USA.
authors: Jie
authors: J
authors: Wang
authors: 1. The Virginia G. Piper Center for Personalized Diagnostics, Biodesign Institute, Arizona State University, Tempe, AZ 85287, USA.
authors: Ji
authors: J
authors: Qiu
authors: 2. Department of Biological Sciences, University of Pittsburgh, Pennsylvania 15260, USA.
authors: James M
authors: JM
authors: Pipas
authors: 1. The Virginia G. Piper Center for Personalized Diagnostics, Biodesign Institute, Arizona State University, Tempe, AZ 85287, USA.
authors: Joshua
authors: J
authors: LaBaer
mesh_terms: D003001:Cloning, Molecular / Q000379:methods*; D005814:Genes, Viral / Q000235:genetics; D006367:HeLa Cells; D054884:Host-Pathogen Interactions / Q000235:genetics*; D006801:Humans; D016366:Open Reading Frames / Q000235:genetics; D016133:Polymerase Chain Reaction; D020543:Proteome / Q000235:genetics* / Q000378:metabolism; D012412:Rubella virus / Q000235:genetics / Q000502:physiology; D014764:Viral Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism
chemical_list: D020543:Proteome; D014764:Viral Proteins
keywords: Gateway® cloning; Gene synthesis; Nucleic Acid Programmable Protein Arrays (NAPPA).; ORFeome; Panviral proteome; Virus-host interactions