表达鸡传染性法氏囊病病毒VP2基因的重组火鸡疱疹病毒的构建及生物学特性分析

doi:10.11843/j.issn.0366-6964.2023.02.022

Abstract

Abstract: Infectious bursal disease (IBD) is a highly lethal and immunosuppressive infectious disease that seriously harms the poultry industry. In order to develop a recombinant herpesvirus of turkey (HVT) vector vaccine against IBD, a recombinant HVT expressing the protective antigen VP2 gene of infectious bursal disease virus (IBDV) was constructed and its biological characteristics in vitro were analyzed. The VP2 gene of very virulent IBDV (vvIBDV) was amplified by RT-PCR and cloned into pCI vector to obtain the recombinant eukaryotic expression plasmid pCI-VP2. The expression frame of VP2 gene carrying CMV promoter was cut off by restriction endonuclease and ligated into entry plasmid pENTR to construct a recombinant entry plasmid pENTR-VP2. The recombinant expression fosmid H3-VP2 were constructed by LR recombination reaction between pENTR-VP2 entry plasmid and HVT recombinant fosmid H3-Kan/ccdB. The recombinant virus rHVT-VP2 was obtained by co-transfection of H3-VP2 with four other fosmids that overlap each other and cover the whole genome of HVT into chicken embryo fibroblasts (CEF). After continuously passaged in CEF for 20 generations, the recombinant virus was detected by PCR, indirect immunofluorescence, and Western blot. The growth curve of recombinant virus in vitro was determined and its replication characteristics in vitro were analyzed. The results showed that the recombinant virus rHVT-VP2 stably expressed VP2 protein, and the replication ability of rHVT-VP2 in CEF was not significantly different from that of the parent virus. The recombinant virus rHVT-VP2 induced virus neutralization antibodies against IBDV and provided 90% protection against vvIBDV-induced death in chickens. The construction of recombinant virus rHVT-VP2 lays a foundation for the development of recombinant HVT live vector vaccine against IBD and is of great significance for the prevention and control of this disease.

Key words: infectious bursal disease, herpesvirus of turkey, VP2 gene, biological characteristics

CLC Number:

S852.659.4

LI Xiaohan, YANG Fuchun, LIU Rui, GAO Li, CUI Hongyu, ZHANG Yanping, LIU Changjun, QI Xiaole, WANG Xiaomei, GAO Yulong, LI Kai. Construction of Recombinant Herpesvirus of Turkey Expressing Infectious Bursal Disease Virus VP2 Gene and Analysis of Its Biological Characteristics[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(2): 656-662.

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Construction of Recombinant Herpesvirus of Turkey Expressing Infectious Bursal Disease Virus VP2 Gene and Analysis of Its Biological Characteristics

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