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中文摘要：
为了解日本血吸虫Wnt信号通路受体和配体间的相互作用关系,构建了日本血吸虫5个Wnt配体和4个Frizzled受体的真核表达载体,并转染293T细胞进行了表达。间接免疫荧光试验结果显示,所构建的p3×FLAG-Wnts、pRK5-frizzleds-myc真核表达载体均能在293T细胞中被成功表达。Western-blot结果显示,所表达的重组蛋白的分子质量与理论分子质量相符。双重间接免疫荧光试验结果显示,两组共转染受体和配体重组质粒p3×FLAG-Wnt2、p3×FLAG-Wnt4与pRK5-frizzled5-myc、pRK5-frizzled8-myc能够在同一细胞内共表达。本研究为下一步分析Wnt受体和配体间相互作用关系奠定了试验基础。
英文摘要：
In order to understand the specificity of ligand-receptor interaction of Wnt signaling pathway of Schistosoma japonicum,the eukaryotic expression vectors of Wnt and frizzled genes were constructed and transformed into 293 T cells.The expression results were examined by indirect immunofluorescence assay and Westernblotting.All recombinant plasmids were successfully expressed in 293 T cells as observed by indirect immunofluorescence assay.The molecular weight of recombinant protein was coincident with the theoretical molecular weight as determined by Western-blotting.In addition,two group co-transfected plasmids p3 ×FLAG-Wnt2/p3 ×FLAG-Wnt4 and pRK5-frizzled 5-myc/pRK5-frizzled 8-myc were also expressed in the same 293 T cells as visualized in double immunofluorescence.These results provided the preliminary basis for analyzing the specific interactions between Wnt and Frizzled.
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