羊口疮病毒F1L、B2L基因重组腺病毒构建及对小鼠的免疫效果分析

doi:10.11843/j.issn.0366-6964.2018.08.015

Abstract

Abstract:

The recombinant adenoviruses of orf virus (ORFV) was constructed by splicing F1L and B2L genes with the self-cut T2A and P2A peptide, and its immune effects were evaluated, which provided the basis for vaccine development of ORFV. The target genes of F1L and B2L from ORFV-F416 strain were amplified by PCR, and the recombinant adenovirus shuttle plasmid was constructed. The shuttle plasmid and backbone plasmid were co-transfected into HEK-293 cells and the ORFV recombinant adenovirus was packaged. The recombinant adenovirus was used to infect MDBK cells and the supernatant was collected to immunize ICR mice. Indirect ELISA, lymphocyte proliferation and ORFV challenge were conducted to evaluate the immunogenicity of mice. The results showed that pAV-CMV-F1L-T2A-B2L-P2A-EGFP recombinant adenovirus was successfully obtained, and its TCID₅₀ was 10^-5.375·mL^-1. PCR and IFA results showed that the target gene was expressed both in gene and protein levels. After immunization with ORFV recombinant adenovirus, the antibody level of mice increased rapidly. ORFV recombinant adenovirus group showed much higher ability of T lymphocyte proliferation, which was significantly higher than that of PBS group (P<0.01). The results of this article showed that the recombinant adenovirus of F1L and B2L genes had positive protective effect in mice, which could stimulate humoral and cellular immune responses, and have no effects on feed intake.

CLC Number:

S852.659.1

CHEN Xiao, GE Lei, DAI Jian-jun, LI Li, ZHANG De-fu, WU Cai-feng, ZHANG Shu-shan. Construction of Recombinant Adenovirus with Orf Virus F1L and B2L Genes and Analysis of Its Immune Efficacy in Mice[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(8): 1701-1708.

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Construction of Recombinant Adenovirus with Orf Virus F1L and B2L Genes and Analysis of Its Immune Efficacy in Mice

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