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ORIGINAL RESEARCH article

Front. Microbiol.
Sec. Virology
Volume 15 - 2024 | doi: 10.3389/fmicb.2024.1442551

A novel approach for efficient co-expressing two foreign genes based on the reverse genetic system of Newcastle disease virus

Provisionally accepted
Ting Lan Ting Lan 1,2*Qilong Liu Qilong Liu 1,2Jinying Ge Jinying Ge 1*Yong Wang Yong Wang 2*
  • 1 State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute (CAAS), Harbin, Heilongjiang Province, China
  • 2 Harbin Medical University, Harbin, China

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

    Newcastle disease virus (NDV) is an ideal model for exploring the mechanisms of the virus, it is also an optimal vector for develop vector vaccines and for cancer therapy. A reverse genetic system of NDV mukteswar strain controlled by eukaryotic cellular RNA polymerase II promoter was established by reverse genetics technology. Based on the reverse genetic system, an open reading frame of the enhanced green fluorescent protein (EGFP) gene be inserted between the P and M genes of the viral genome, and flanked with the gene start (GS) sequence and gene end (GE) sequence to form an independent transcription unit. The rescued virus was amplified in specific-pathogen-free (SPF) chicken embryos for 10 generations, and the results showed that the recombinant virus could stably express the exogenous gene for at least 10 generations. Efficient expression of two exogenous genes synchronous is essential for the development of NDV-based multivalent vaccine candidates. Explore the possibility of simultaneous and efficient expression of two exogenous genes based on NDV vector, in the present study, a recombinant virus with co-expression of EGFP and cherry fluorescent protein (CFP) that be inserted between the intergenic region of P/M gene as two independent transcription units be succeeded rescued. The results showed that the two exogenous genes could be expressed synchronously and efficiently. The results of biological analysis of the expression efficiency of exogenous genes showed that the EGFP in recombinant viruses with two exogenous genes was slightly lower than that of recombinant viruses with one exogenous gene, but the expression efficiency of CFP in recombinant viruses with two exogenous genes was higher than EGFP in both viruses. These recombinant viruses have similar growth kinetics but with a little attenuated in virulence compared with parental virus. In conclusion, these data indicated that this study successfully established the reverse genetic system of NDV Mukteswar strain, and achieved the purpose of efficient expression of two exogenous genes synchronously in a novel approach, laying the foundation for the development of multivalent vaccines or tumor therapeutics using NDV as a vector.

    Keywords: Newcastle disease virus, reverse genetic system, Recombinant virus, Foreign gene, 32 coexpression 33 34

    Received: 02 Jun 2024; Accepted: 12 Nov 2024.

    Copyright: © 2024 Lan, Liu, Ge and Wang. 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:
    Ting Lan, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute (CAAS), Harbin, 150001, Heilongjiang Province, China
    Jinying Ge, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute (CAAS), Harbin, 150001, Heilongjiang Province, China
    Yong Wang, Harbin Medical University, Harbin, China

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