猪附红细胞体eno基因重组蛋白的制备及免疫效果研究

doi:10.16431/j.cnki.1671-7236.2020.07.029

Abstract

Abstract: In order to study the immune effect of the recombinant protein of Mycoplasma suis eno gene,the Mycoplasma suis eno gene was linked with pET-15b vector and transformed into E.coli BL21 competent cells.The recombinant Mycoplasma suis eno protein (named rMseno) was used to immunize mice,and the immune response of the recombinant protein in mice was evaluated.Twenty four-week-old male Kunming mice were randomly divided into four groups:Group A,group B,group C and group D.Immune group A was inoculated with recombinant protein rMseno.Immune group B was inoculated with E.coli-Mseno induced by IPTG.Control group C was inoculated with the same amount of PBS.Control group D was inoculated with E.coli-Mseno which was not induced expression.The levels of specific antibody against Mycoplasma suis,IFN-γ and IL-4 cytokines in serum were determined by ELISA,and the proliferation of T lymphocyte in mice was detected by spleen lymphocyte test,the experimental data were analyzed statistically.The results showed that the target gene fragment of the recombinant pET-15b-eno plasmid was 1 632 bp,and the purified recombinant protein rMseno was 61 ku,which could be recognized by the mouse anti-Mycoplasma suis serum.The levels of anti-Mycoplasma suis specific antibody,IFN-γ,IL-4 cytokines and T lymphocyte proliferation index in the sera of mice in immune group A and immune group B were significantly or extremely significantly higher than those in control groups (P<0.05;P<0.01).The results showed that eno gene recombinant protein of Mycoplasma suis could induce high humoral immunity and cellular immune response in mice.

Key words: Mycoplasma suis; purification of recombinant protein; Kunming mice; immune effect

CLC Number:

S858.28

YAN Zongbin, WU Shengjun, YANG Jing, YAN Kexin, XIANG Siyu, XU Yingtian, XUE Shujiang, CHAI Fanghong. Preparation of Recombinant Protein of Mycoplasma suis eno Gene and Its Immunity Effect[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(7): 2231-2238.

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Preparation of Recombinant Protein of Mycoplasma suis eno Gene and Its Immunity Effect

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