Construction and immune protection evaluation of recombinant virus expressing Newcastle disease virus F protein by the l
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ORIGINAL PAPER
Construction and immune protection evaluation of recombinant virus expressing Newcastle disease virus F protein by the largest intergenic region of fowlpox virus NX10 Yan Zhao1,2 · Zongxi Han2 · Xiaocai Zhang2 · Xuemei Zhang2 · Junfeng Sun2 · Deying Ma1 · Shengwang Liu2 Received: 30 March 2020 / Accepted: 26 September 2020 © Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract Fowlpox virus (FPV) is used as a vaccine vector to prevent diseases in poultry and mammals. The insertion site is considered as one of the main factors influencing foreign gene expression. Therefore, the identification of insertion sites that can stably and efficiently express foreign genes is crucial for the construction of recombinant vaccines. In this study, we found that the insertion of foreign genes into ORF054 and the ORF161/ORF162 intergenic region of the FPV genome did not affect replication, and that the foreign genes inserted into the intergenic region were more efficiently expressed than when they were inserted into a gene. Based on these results, the recombinant virus rFPVNX10-NDV F–E was constructed and immune protection against virulent FPV and Newcastle disease virus (NDV) was evaluated. Tests for anti-FPV antibodies in the vaccinated chickens were positive within 14 days post-vaccination. After challenge with FPV102, no clinical signs of FP were observed in vaccinated chickens, as compared to that in the control group (unvaccinated), which showed 100% morbidity. Low levels of NDV-specific neutralizing antibodies were detected in vaccinated chickens before challenge. After challenge with NDV ck/CH/LHLJ/01/06, all control chickens died within 4 days post-challenge, whereas 5/15 vaccinated chickens died between 4 and 12 days post-challenge. Vaccination provided an immune protection rate of 66.7%, whereas the control group showed 100% mortality. These results indicate that the ORF161/ORF162 intergenic region of FPVNX10 can be used as a recombination site for foreign gene expression in vivo and in vitro. Keywords Fowlpox virus NX10 strain · Non-essential sites · Intergenic region · Gaussia luciferase · Enhanced green fluorescent protein · Immune protection
Introduction Edited by Joachim Jakob Bugert. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s11262-020-01799-5) contains supplementary material, which is available to authorized users.
Fowlpox virus (FPV) is the prototype member of the Avipoxvirus genus in the Chordopoxvirinae subfamily of the family Poxviridae according to the International Committee
* Deying Ma [email protected]
Junfeng Sun [email protected]
* Shengwang Liu [email protected]
1
College of Animal Science and Technology, Northeast Agricultural University, No. 600 Changjiang Road, Xiangfang District, Harbin 150030, China
2
Division of Avian Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, The Chinese Academy of Agricultural Sciences, Xiangf
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