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当前位置: 首页 > 文章 > 锡兰钩虫谷胱甘肽-S-转移酶基因的克隆表达与免疫原性分析 中国兽医学报 2020 (7) 1361-1365
Position: Home > Articles > Cloning and expression of Ancylostoma ceylanicum GST gene and its immunogenicity analysis Chinese Journal of Veterinary Science 2020 (7) 1361-1365

锡兰钩虫谷胱甘肽-S-转移酶基因的克隆表达与免疫原性分析

作  者：
杭建雄;黄岳;颜欣欣;刘云秋;赵琪;何龙;冉荣坤;李秀;刘菊枚;李国清
单  位：
华南农业大学兽医学院
关键词：
锡兰钩虫;谷胱甘肽-S-转移酶;基因克隆;原核表达;免疫原性
摘  要：
为了克隆表达锡兰钩虫谷胱甘肽-S-转移酶(Ace-GST)基因,制备多克隆抗体并分析其免疫原性,为疫苗候选分子的筛选奠定基础。根据GenBank上锡兰钩虫GST基因序列设计引物,利用RT-PCR法扩增Ace-GST基因,将扩增产物克隆至pET-32a载体并转入E.coli BL21(DE3)宿主菌表达,重组蛋白纯化后免疫昆明小鼠,分别用间接ELISA法和MTT法检测免疫鼠血清IgG抗体水平与脾淋巴细胞增殖情况。结果显示Ace-GST基因ORF全长468 bp,编码155个氨基酸,与ν类GSTs聚为同一分支;重组Ace-GST相对分子质量为36 000,可诱导产生特异性IgG抗体,并能显著刺激免疫小鼠脾淋巴细胞增殖。结果表明重组Ace-GST蛋白能诱导昆明小鼠产生特异性免疫应答,具有良好的免疫原性。
译  名：
Cloning and expression of Ancylostoma ceylanicum GST gene and its immunogenicity analysis
作  者：
HANG Jian-xiong;HUANG Yue;YAN Xin-xin;LIU Yun-qiu;ZHAO Qi;HE Long;RAN Rong-kun;LI xiu;LIU Ju-mei;LI Guo-qing;College of Veterinary Medicine,South China Agricultural University;
关键词：
Ancylostoma ceylanicum;;glutathione-S-transferase(GSTs);;gene cloning;;prokaryotic expression;;immunogenicity
摘  要：
This study aims at cloning and expressing the glutathione-S-transferase gene from Ancylostoma ceylanicum(Ace-GST),and analyzing its immunogenicity for screening vaccine candidate against A.ceylanicum.The Ace-GST gene was amplified by RT-PCR method using primers designed based on the coding sequence of A.ceylanicum GST in GenBank.The amplified product was cloned into pET-32 a vector and transferred into E.coli BL21(DE3) for expression.Kunming mice were immunized with the recombinant protein after purification,and the levels of serum IgG antibody and spleen lymphocyte proliferation were detected by indirect ELISA and MTT assay,respectively.The results showed that the Ace-GST gene had a full-length of 468 bp,encoding 155 amino acids,and belonging to ν class GSTs.The recombinant Ace-GST had a molecular weight of 36 000,which could induce the production of specific IgG antibody,and significantly stimulated the proliferation of spleen lymphocytes in the immunized mice.It is concluded that recombinant Ace-GST protein can induce specific immune response in Kunming mice,indicating good immunogenicity.

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