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利用鸭肠炎病毒载体进行鹅星状病毒Cap蛋白的体外表达。

In vitro expression of the goose astrovirus Cap protein delivered with a duck enteritis virus vector.

作者信息

Chen Liu, Zhu Yinchu, Yun Tao, Ye Weicheng, Ni Zheng, Hua Jionggang, Fu Yuan, Zhang Cun

机构信息

State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro- Products, Institute of Animal Husbandry and Veterinary Sciences, Zhejiang Academy of Agricultural Sciences, Hangzhou, China.

出版信息

BMC Vet Res. 2025 May 2;21(1):311. doi: 10.1186/s12917-025-04654-7.

DOI:10.1186/s12917-025-04654-7
PMID:40317044
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC12046716/
Abstract

BACKGROUND

Goose astrovirus (GAstV) is an emerging pathogen that is widely distributed throughout China and can cause visceral gout, resulting in serious economic losses for the goose industry. Open reading frame 2 (ORF2) of this virus encodes the precursor capsid protein, which is essential for the assembly and antigenicity of these virions. To construct a bi-valent vaccine for controlling GAstV and duck enteritis virus (DEV) infection, an infectious bacterial artificial chromosome (BAC) clone of the DEV vaccine strain pDEV-EF1 was used to establish a recombinant DEV vector for GAstV ORF2 gene delivery.

METHODS

GAstV ORF2 expression frame was inserted into the US7 and US8 intergenic region of DEV genome by Red E/T two-step recombinant technology, then the recombinant virus rDEV-GAstV ORF2 was rescued by transfecting recombinant clone pDEV-GAstV ORF2 into chicken embryonic fibroblasts (CEFs). The expression of ORF2 in CEFs and formation of virus-like particles (VLPs) were analysed by Western blotting, indirect immunofluorescence assay (IFA) and immunogold electron microscopy (IEM), individually. And protein celluar localization was analysed by IFA.

RESULTS

Using this rDEV-GAstV ORF2 vector to infect CEFs was sufficient to elicit GAstV Cap protein expression, as confirmed by Western blotting and IFA. IEM also revealed the formation of VLPs within cells expressing this Cap protein.

CONCLUSIONS

DEV is a good viral vector for GAstV ORF2 gene delivery and these results provide a basis for the development of a bivalent vaccine for controlling DEV and GAstV infections.

摘要

背景

鹅星状病毒（GAstV）是一种新出现的病原体，在中国广泛分布，可引起内脏痛风，给养鹅业造成严重经济损失。该病毒的开放阅读框2（ORF2）编码前体衣壳蛋白，这对于这些病毒粒子的组装和抗原性至关重要。为构建用于防控GAstV和鸭肠炎病毒（DEV）感染的二价疫苗，利用DEV疫苗株pDEV-EF1的感染性细菌人工染色体（BAC）克隆建立了用于递送GAstV ORF2基因的重组DEV载体。

方法

通过Red E/T两步重组技术将GAstV ORF2表达框插入DEV基因组的US7和US8基因间隔区，然后将重组克隆pDEV-GAstV ORF2转染至鸡胚成纤维细胞（CEF）中拯救重组病毒rDEV-GAstV ORF2。分别通过蛋白质免疫印迹法、间接免疫荧光试验（IFA）和免疫金电子显微镜（IEM）分析ORF2在CEF中的表达及病毒样颗粒（VLP）的形成。并通过IFA分析蛋白的细胞定位。

结果

蛋白质免疫印迹法和IFA证实，使用该rDEV-GAstV ORF2载体感染CEF足以诱导GAstV Cap蛋白表达。IEM还揭示了在表达该Cap蛋白的细胞内形成了VLP。

结论

DEV是用于递送GAstV ORF2基因的良好病毒载体，这些结果为开发用于防控DEV和GAstV感染的二价疫苗奠定了基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7424/12046716/c1bf4bfef122/12917_2025_4654_Fig9_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7424/12046716/c2d6e766529b/12917_2025_4654_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7424/12046716/f07020fab2db/12917_2025_4654_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7424/12046716/79715005c47c/12917_2025_4654_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7424/12046716/b0388f830cb1/12917_2025_4654_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7424/12046716/e8252d4cca44/12917_2025_4654_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7424/12046716/381208c5d61c/12917_2025_4654_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7424/12046716/bbe698f43c26/12917_2025_4654_Fig7_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7424/12046716/ddb72eae00a3/12917_2025_4654_Fig8_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7424/12046716/c1bf4bfef122/12917_2025_4654_Fig9_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7424/12046716/c2d6e766529b/12917_2025_4654_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7424/12046716/f07020fab2db/12917_2025_4654_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7424/12046716/79715005c47c/12917_2025_4654_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7424/12046716/b0388f830cb1/12917_2025_4654_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7424/12046716/e8252d4cca44/12917_2025_4654_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7424/12046716/381208c5d61c/12917_2025_4654_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7424/12046716/bbe698f43c26/12917_2025_4654_Fig7_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7424/12046716/ddb72eae00a3/12917_2025_4654_Fig8_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7424/12046716/c1bf4bfef122/12917_2025_4654_Fig9_HTML.jpg

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Recombinant duck enteritis virus bearing the hemagglutinin genes of H5 and H7 influenza viruses is an ideal multivalent live vaccine in ducks.
携带 H5 和 H7 流感病毒血凝素基因的重组鸭肠炎病毒是鸭子理想的多价活疫苗。
Emerg Microbes Infect. 2024 Dec;13(1):2284301. doi: 10.1080/22221751.2023.2284301. Epub 2024 Jan 4.
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