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ORIGINAL RESEARCH article

Front. Vet. Sci.
Sec. Veterinary Infectious Diseases
Volume 11 - 2024 | doi: 10.3389/fvets.2024.1418760

Rescuing Newcastle Disease Virus with Tag for Screening Viral-Host Interacting Proteins Based on Highly Efficient Reverse Genetics

Provisionally accepted
Ruiwei Wang Ruiwei Wang Xuhong Cao Xuhong Cao Kejia Lu Kejia Lu Zhengwu Chang Zhengwu Chang Xiaoyu Dong Xiaoyu Dong Hanwei Guo Hanwei Guo Xi Wei Xi Wei Ruyi Dang Ruyi Dang Juan Wang Juan Wang Xinglong Wang Xinglong Wang Sa Xiao Sa Xiao Haijin Liu Haijin Liu Zengqi Yang Zengqi Yang *
  • College of Veterinary Medicine, Northwest A&F University, Xianyang, China

The final, formatted version of the article will be published soon.

    The interaction between viral proteins and host proteins plays a crucial role in the process of virus infecting cells. Tags such as HA, His, and Flag do not interfere with the function of fusion proteins and are commonly used to study protein-protein interactions. Adding these tags to viral proteins will address the challenge of the lack of antibodies for screening host proteins that interact with viral proteins during infection.Obtaining viruses with tagged fusion proteins is crucial. This study established a new reverse genetic system with T7 promoter and three plasmids, which efficiently rescued Newcastle disease virus (NDV) regardless of its ability to replicate in cells.Subsequently, using this system, NDV containing a HA-tagged structural protein and NDV carrying a unique tag on each structural protein were successfully rescued. These tagged viruses replicated normally and exhibited genetic stability. Based on tag antibodies, every NDV structural protein was readily detected and showed correct subcellular localization in infected cells. After infecting cells with NDV carrying HAtagged M protein, several proteins interacting with the M protein during the infection process were screened using HA tag antibodies. The establishment of this system laid the foundation for comprehensive exploration of the interaction between NDV proteins and host proteins.

    Keywords: Newcastle disease virus, Reverse Genetics, protein tag, M protein, Viral-host interaction

    Received: 17 Apr 2024; Accepted: 04 Jul 2024.

    Copyright: © 2024 Wang, Cao, Lu, Chang, Dong, Guo, Wei, Dang, Wang, Wang, Xiao, Liu and Yang. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

    * Correspondence: Zengqi Yang, College of Veterinary Medicine, Northwest A&F University, Xianyang, China

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