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伤寒沙门菌鞭毛素基因fljB:z66的克隆表达及其多克隆抗体的制备 被引量：3

Cloning,expression and polyclonal antibody preparation of flagellin gene fljB:z66 of Salmonella enterica serover Typhi
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摘要 目的:克隆表达H:z66阳性伤寒沙门菌鞭毛素基因fljB:z66,并纯化制备相应的多克隆抗体。方法:利用PCR技术从H:z66阳性伤寒沙门菌基因组DNA中得到鞭毛素基因fljB:z66,将该基因克隆到表达载体pET-28 a(+)上并在大肠埃希菌JM109中进行表达;fljB:z66的表达产物经N i柱纯化后作为抗原免疫兔以制备多克隆抗体。结果:经PCR及序列分析证实,伤寒沙门菌鞭毛素基因fljB:z66被成功导入表达载体pET-28 a(+),并在大肠埃希菌JM109中获得了高效表达,以此作为抗原成功制备了兔抗z66的多克隆抗体。结论:成功克隆表达了H:z66阳性伤寒沙门菌鞭毛素基因fljB:z66,并制备了相应的多克隆抗体,为今后深入研究H:z66阳性伤寒沙门菌的单向相变换机制奠定了基础。 Objective： To clone and express the flagellin gene fljB：z66 of H：z66 positive Salmonella enterica serover Typhi,and prepare the anti-z66 polyclonal antibodies.Methods： Specific primers were designed to amplify the flagellin gene fljB：z66 from the chromosome DNA of H：z66 positive Salmonella enterica serover Typhi GIFU10007.The amplicon was inserted into the expression vector pET-28a（＋）,which was then transferred to E.coli JM109 to be expressed.The recombinant protein FljB-His6 was purified with Ni-TED packed column and was used as immunogen to prepare the polyclonal antibody. Results： PCR andsequencing analysis demonstrated that the flagellin gene fljB：z66 of H：z66 positive Salmonella enterica serover Typhi was inserted into the vector pET-28a（ ＋ ） and was expressed highly in E. coli JM109. The rabbit anti-z66 polyclonal antibody was prepared successfully. Conclusion： This study was a foundation to research the mechanism underlying the the unidirectional flagellar phase variation of H：z66 positive Salmonella enterica serover Typhi.
作者 张海方 高宇琳 黄新祥 生秀梅 徐顺高 许化溪
机构地区 江苏大学基础医学与医学技术学院医学生物化学系 东莞市妇幼保健院检验科
出处 《江苏大学学报（医学版）》 CAS 2009年第5期376-379,共4页 Journal of Jiangsu University：Medicine Edition
基金 国家自然科学基金资助项目(30870095)
关键词 伤寒沙门菌 fljB:z66 多克隆抗体 Salmonella enterica serover Typhi fljB：z66 polyclonal antibody
分类号 R392 [医药卫生—免疫学]
  • 相关文献

参考文献12

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二级参考文献9

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同被引文献37

  • 1茅凌翔,朱超望,许化溪,黄新祥.伤寒沙门菌phoP基因缺陷变异株的制备[J].江苏大学学报（医学版）,2007,17(2):145-149. 被引量：15
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引证文献3

江苏大学学报（医学版）
2009年 第5期

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