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目的 :构建人成熟型TGF - β1基因原核表达载体 ,表达并纯化人成熟型TGF - β1融合蛋白。方法 :应用基因重组的方法 ,构建成熟型TGF - β1原核表达载体 ,诱导表达人成熟型TGF - β1的融合蛋白 ,并应用亲和层析法纯化重组融合蛋白 ,应用Western -Blot鉴定其抗原性。结果 :成功构建人成熟型TGF - β1原核表达载体 ,获得人成熟型TGF - β1融合蛋白 ,其抗原性较好。结论 :人成熟型TGF - β1基因原核表达载体的成功构建 ,可高效表达人成熟型TGF - β1融合蛋白 ,为进一步研究其功能和获得其多克隆抗体奠定了基础 ,并为开发国产人TGF - β1诊断试剂做必要的准备 ,对肝纤维化的防治具有较大意义。
Objective: To construct prokaryotic expression vector of human mature TGF - β1 gene and to express and purify human mature TGF - β1 fusion protein. Methods: The prokaryotic expression vector of mature TGF - β1 gene was constructed by gene recombination. The fusion protein of human mature TGF - β1 was induced. The recombinant fusion protein was purified by affinity chromatography and its antigenicity was identified by Western - blot . Results: The prokaryotic expression vector of human mature TGF - β1 was successfully constructed and the human mature TGF - β1 fusion protein was obtained. The antigenicity was good. Conclusion: The successful construction of human mature TGF - β1 gene prokaryotic expression vector can efficiently express human mature TGF - β1 fusion protein, which lays the foundation for further study on its function and obtaining its polyclonal antibody. β1 diagnostic reagents to make the necessary preparations for the prevention and treatment of liver fibrosis has great significance.