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Viruses 2014, 6(6), 2428-2443; https://doi.org/10.3390/v6062428

Efficient Strategy to Generate a Vectored Duck Enteritis Virus Delivering Envelope of Duck Tembusu Virus

1,2,3
,
1,2,3,4
and
1,2,3,*
1
State Key Laboratory of Agricultural Microbiology, Huazhong Agricultura University, Wuhan 430070, China
2
Key Laboratory of Development of Veterinary Diagnostic Products, Ministry of Agriculture, Wuhan 430070, China
3
College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China
4
College of Life Science and Technology, AnQing Normal University, AnQing 246011, China
*
Author to whom correspondence should be addressed.
Received: 10 March 2014 / Revised: 18 May 2014 / Accepted: 21 May 2014 / Published: 20 June 2014
(This article belongs to the Special Issue Virus-based Vaccines)
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Abstract

Duck Tembusu virus (DTMUV) is a recently emerging pathogenic flavivirus that has resulted in a huge economic loss in the duck industry. However, no vaccine is currently available to control this pathogen. Consequently, a practical strategy to construct a vaccine against this pathogen should be determined. In this study, duck enteritis virus (DEV) was examined as a candidate vaccine vector to deliver the envelope (E) of DTMUV. A modified mini-F vector was inserted into the SORF3 and US2 gene junctions of the attenuated DEV vaccine strain C-KCE genome to generate an infectious bacterial artificial chromosome (BAC) of C-KCE (vBAC-C-KCE). The envelope (E) gene of DTMUV was inserted into the C-KCE genome through the mating-assisted genetically integrated cloning (MAGIC) strategy, resulting in the recombinant vector, pBAC-C-KCE-E. A bivalent vaccine C-KCE-E was generated by eliminating the BAC backbone. Immunofluorescence and western blot analysis results indicated that the E proteins were vigorously expressed in C-KCE-E-infected chicken embryo fibroblasts (CEFs). Duck experiments demonstrated that the insertion of the E gene did not alter the protective efficacy of C-KCE. Moreover, C-KCE-E-immunized ducks induced neutralization antibodies against DTMUV. These results demonstrated, for the first time, that recombinant C-KCE-E can serve as a potential bivalent vaccine against DEV and DTMUV. View Full-Text
Keywords: duck enteritis virus; bacterial artificial chromosome; mating-assisted genetically integrated cloning; DTMUV: bivalent vaccine duck enteritis virus; bacterial artificial chromosome; mating-assisted genetically integrated cloning; DTMUV: bivalent vaccine
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Zou, Z.; Liu, Z.; Jin, M. Efficient Strategy to Generate a Vectored Duck Enteritis Virus Delivering Envelope of Duck Tembusu Virus. Viruses 2014, 6, 2428-2443.

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