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




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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.



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





Autor: Zhong Zou 1,2,3, Zhigang Liu 1,2,3,4 and Meilin Jin 1,2,3,*

Fuente: http://mdpi.com/



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