| 摘要: |
| 鸭肠炎病毒(Duck enteritis virus, DEV),又称为鸭瘟病毒,是一种只感染雁形目禽类的疱疹病毒。DEV具有基因组大、非必需基因多、能插入外源基因的容量大、遗传稳定等优点,其庞大而复杂的基因组为外源基因提供了诸多可插入位点,以DEV为载体,成功表达了禽流感病毒HA蛋白[1]、鸭病毒性肝炎病毒VP0蛋白[2]、鸭坦布苏病毒E基因[3]以及鹅细小病毒VP2蛋白[4]。构建重组DEV的重要一步是将报告基因插入到基因组中,目前常用的报告基因有绿色荧光蛋白(GFP)、增强型绿色荧光蛋白(EGFP)、红色荧光蛋白(RFP)以及LacZ报告基团。本研究用RFP报告基团插入DEV UL2基因中,获得表达红色荧光的重组病毒,一步生长曲线表明,RFP对DEV的生长无影响；连续传代12代,RFP能够稳定表达,为DEV载体研究奠定基础。 |
| 关键词: 鸭肠炎病毒 红色荧光蛋白 重组病毒 |
| DOI： |
| 投稿时间：2021-06-11修订日期：2021-09-21 |
| 基金项目:国家重点研发计划“水禽重要疫病免疫防控新技术研究”(2017YFD0500800) |
|
| Construction and Characterization of Recombinant Duck Enteritis Virus Expressing the Red Flourescent Protein |
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| (China Institute of Veterinary Drug Control,Beijing,100081) |
| Abstract: |
| Duck enteritis virus (DEV), also known as duck plague virus, is a herpes virus that only infects birds of Anseriformes. It has the advantages of large genome, many non-essential genes, large capacity for inserting foreign genes, genetic stability, etc. Its large and complex genome provides many sites for insertion of foreign genes. DEV is used as vectors for the delivery and in vivo expression of antigens of pathogens, such as avian Influenza virus HA protein, duck viral hepatitis virus VP1 protein, duck tambusu virus E gene, and goose parvovirus VP2 protein. An important step in constructing a recombinant DEV is to insert a reporter gene into the genome. Currently commonly used reporter genes are green fluorescent protein (GFP), enhanced green fluorescent protein (EGFP), red fluorescent protein (RFP) and LacZ. In this study, the RFP gene was inserted into the DEV UL2 gene to obtain a recombinant virus expressing red fluorescence. The one-step growth curve showed that RFP has no effect on the growth of DEV; RFP was continuously passaged for 12 generations and could be stably expressed. The result contributes to construct DEV-vectored vaccines. |
| Key words: Duck enteritis virus RFP recombinant virus |
