赖型钩体毒力相关基因ompL17的克隆表达及产物的体内分布

来源 :现代预防医学 | 被引量 : 0次 | 上传用户:kevil2009
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[目的]观察ompL17基因表达产物的免疫原性以及该表达蛋白在动物模型中的分布情况。[方法]提取钩端螺旋体017株基因组DNA,扩增出ompL17基因,与表达载体pGEX-4T-1重组,诱导表达OMPL17蛋白,免疫动物获得相应抗体,通过免疫组织化学技术观察该蛋白在动物模型组织当中的分布。[结果]PCR和酶切分析证实构建成功了表达载体,SDS-PAGE和Western-blot分析证实诱导表达出OMPL17蛋白,通过免疫组织化学技术观察到了该蛋白在动物模型中的分布情况。[结论]成功表达出ompL17基因的蛋白,观察到豚鼠动物模型中该蛋白的分布情况,为赖型钩体的分子机制的阐明奠定了基础。 [Objective] To observe the immunogenicity of ompL17 gene expression product and the distribution of the expressed protein in animal model. [Method] The genomic DNA of 017 strain of Leptospira was extracted and the ompL17 gene was amplified. The recombinant plasmid was transformed into pGEX-4T-1 vector to express OMPL17 protein. The corresponding antibodies were obtained from the animals. The expression of ompL17 gene was observed by immunohistochemistry in animal models Distribution among organizations. [Result] PCR and restriction analysis confirmed that the constructed expression vector was successfully constructed. The expression of OMPL17 protein was confirmed by SDS-PAGE and Western-blot analysis. The distribution of OMPL17 protein was observed by immunohistochemistry in animal models. [Conclusion] The protein of ompL17 gene was successfully expressed and the distribution of the protein in guinea pig animal model was observed, which laid the foundation for the elucidation of the molecular mechanism of the leptospira.
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