表达猪附红细胞体ENO基因的质粒DNA的构建及免疫效果研究

doi:10.16431/j.cnki.1671-7236.2018.09.028

Abstract

Abstract:

The aim of this experiment was to construct the plasmid DNA expressing ENO gene of Mycoplasma suis and determine its immune effect.The Mycoplasma suis ENO gene was cloned into PVAX1 eukaryotic expression vector,then transfected into Vero cells to express and determine its immune effect.18 BALB/c mice (half male and female) were randomly divided into 3 groups (PVAX1-ENO plasmid DNA immunization group,PVAX1 empty vector control group and PBS control group),6 mice in each group.Mice in each group were immunized with PVAX1-ENO plasmid DNA,PVAX1 empty plasmid and PBS,respectively.Serum was separated and serum ENO protein antibody titers,IgG₁ and IgG_2a antibody levels and IFN-γ and IL-4 cytokine levels were determined by ELISA.Two weeks after the third immunization,3 mice from each group were selected to measure the levels of CD4⁺ and CD8⁺.The results showed that the PVAX1-ENO plasmid DNA was successfully constructed and identified by PCR and restriction endonuclease digestion,and it could be successfully expressed in Vero cells.The level of ENO protein antibody,IgG₁ and IgG_2a antibody,IFN-γ and IL-4 cytokines and CD4⁺ and CD8⁺ in PVAX1-ENO plasmid DNA immunized group were significantly higher than those in PVAX1 empty vector control group and PBS control group (P<0.05).The results showed that PVAX1-ENO plasmid DNA could significantly improve the humoral immunity of BALB/c mice.The cellular immune of BALB/c mice was stimulated to some extent.

Key words: Mycoplasma suis; ENO gene; plasmid DNA; cellular immunity; humoral immunity

CLC Number:

S858.28

ZHAO Yun, WU Shengjun, NI Tingting, WANG Miao, ZHANG Shoufa, XU Yingtian, XUE Shujiang. Construction and Immunological Effect Analysis of Plasmid DNA Expressing ENO Gene of Mycoplasma suis[J]. , 2018, 45(9): 2559-2565.

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Construction and Immunological Effect Analysis of Plasmid DNA Expressing ENO Gene of Mycoplasma suis

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