| 分子佐剂鞭毛素对弓形虫SAG1蛋白抗体应答的影响 |
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| 引用本文： | 黄驹辉,龚丽贞,唐静静,黄志坚,殷光文.分子佐剂鞭毛素对弓形虫SAG1蛋白抗体应答的影响[J].福建农林大学学报(自然科学版),2017,46(3). |
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| 作者姓名： | 黄驹辉 龚丽贞 唐静静 黄志坚 殷光文 |
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| 作者单位： | 福建农林大学动物科学学院/福建省动物药物工程实验室,福建 福州,350002 |
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| 基金项目： | 国家自然基金青年项目，福建省自然科学基金面上项目，福建省中青年教师教育科研项目，大学生创新创业训练计划项目 |
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| 摘 要： | 将沙门氏菌鞭毛素和弓形虫免疫优势表面抗原1(SAG1)融合表达,研究其在小鼠上激发的体液免疫应答.首先,采用PCR扩增SAG1片段.其次,将SAG1连接到原核表达载体pET-28a中构建表达质粒pET-28a-SAG1;连接到实验室已有的pET-28a鞭毛素质粒中构建表达质粒pET-28a-F-SAG1;质粒转化至大肠杆菌BL21(DE3)菌株中并进行表达.将纯化后的蛋白免疫小鼠,每隔2周免疫一次,共免疫3次,第3次免疫后15 d取小鼠血清.最后,采用蛋白印迹检测蛋白的大小;用酶联免疫吸附方法检测血清中多克隆抗体的效价.结果表明,诱导出的SAG1蛋白大小为30 ku,F-SAG1大小为70 ku.酶联免疫吸附方法检测的D450 nm均为阴性对照的两倍,表明该多克隆抗体具有较高效价.可见,鞭毛素可以作为分子佐剂促进弓形虫亚单位疫苗的体液免疫应答.
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| 关 键 词： | 弓形虫 表面抗原1(SAG1) 鞭毛素 体液免疫 |
The effect of molecular adjuvant flagellin on the humoral immunity of Toxoplasma gondii induced by subunit vaccine |
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| HUANG Juhui,GONG Lizhen,TANG Jingjing,HUANG Zhijian,YIN Guangwen.The effect of molecular adjuvant flagellin on the humoral immunity of Toxoplasma gondii induced by subunit vaccine[J].Journal of Fujian Agricultural and Forestry University,2017,46(3). |
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| Authors: | HUANG Juhui GONG Lizhen TANG Jingjing HUANG Zhijian YIN Guangwen |
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| Abstract: | Flagellin, a ligand for TLR5, can be an effective adjuvant for vaccine. In this study, we coexpressed the flagellin and Toxoplasma gondii immune dominant antigen SAG1 ( surface antigen 1 ) , the immunogenicity of SAG1 and adjuvant property of flagellin for SAG1 protein in mice were investigated. SAG1 was amplified by PCR, and inserted into prokaryotic expression vector to construct expression plasmid pET-28a-SAG1. Then SAG1 was inserted into pET-28a-F to construct expression plasmid pET-28a-F-SAG1. Subsequently, these 2 plasmids were transformed into BL21 (DE3) E.coli and expressed. Purified proteins were intramuscu-larly injected with the same amount of adjuvant in mice, and immuned at an interval of 2 weeks. After 3 immunization, the serum was obtained and the titer of the polyclonal antibody was evaluated by ELISA. The results showed that SAG1 protein was 30 ku and F-SAG1 was 70 ku. ELISA D450 nm values were twice more than the negative control, indicating a high titer of the polyclonal antibody. In conclusion, flagellin can be an effective adjuvant to promote the humoral immunity of subunit vaccine. |
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| Keywords: | Toxoplasma gondii surface antigen 1 ( SAG1) flagellin humoral immunity |
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