表达牛病毒性腹泻病毒E2蛋白的重组乳酸菌的构建及蛋白免疫原性分析

doi:10.16431/j.cnki.1671-7236.2021.08.030

Abstract

Abstract: The aim of this study was to construct a recombinant Lactococcus lactis expressing E2 antigen protein of Bovine viral diarrhea virus (BVDV), and lay a foundation for further development of oral live vector vaccine of BVDV.The BVDV E2 gene was amplified and sequenced, the gene sequence was optimized and synthesized according to the codon bias of the Lactococcus lactis strain and inserted into the pNZ8148 expression vectors, the recombinant strain pNZ8148-E2/NZ9000 was constructed by electroporation of competent cells of Lactococcus lactis NZ9000, and the induced with 1 ng/mL Nisin, and E2 expression was analyzed by SDS-PAGE and Western blotting.Healthy calves aged 6-12 months were immunized orally with recombinant Lactobacillus pNZ8148-E2/NZ9000, and blood samples were collected and serum was separated at different time after immunization.The antibody level was detected by indirect ELISA.The results showed that the target fragment of 1 149 bp was amplified by PCR.After optimizing the codon preference of Lactococcus lactis, the GC content changed from 45.28% to 34.30%.The pNZ8148-E2 plasmid was identified as the expected size by enzyme digestion.Approximate 42 ku fusion protein was observed from the cell lysates of pNZ8148-E2/NZ9000 in Western blotting.Furthermore, specific anti-E2 IgG was detected in serum of immunized calves.The results suggested oral immunization with E2-expressing Lactobacillus could induce immune response, and the recombinant Lactobacillus had good immunogenicity.

Key words: Bovine viral diarrhea virus (BVDV); E2 gene; Lactococcus lactis; immunogenicity

CLC Number:

S852.65⁺3

WU Tongzhong, NUERSAILIKE Nusufu, HUANG Xin, HAN Mengli, ZHANG Xingxing, ZHANG Qian, WANG Xinhua, HE Yanhua, ZHONG Fagang. Construction of Recombinant Lactococcus Expressing E2 Protein of Bovine Viral Diarrhea Virus and Immunogenicity Analysis of the Protein[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(8): 2975-2981.

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Construction of Recombinant Lactococcus Expressing E2 Protein of Bovine Viral Diarrhea Virus and Immunogenicity Analysis of the Protein

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