【摘要】 目的 用含有人Sy2酪氨酸磷酸酶基因的pOTB7质粒，构建Sy2原核表达重组子，并用该重组子转化原核细胞，表达出含Sy2的融合蛋白，为将来以此融合蛋白为抗原免疫Balb/c小鼠，制备单克隆抗体，进一步研究该基因的功能打好基础。 方法 用PCR扩增Sy2的PP2C结构域序列，限制性内切酶双酶切，构建到表达载体pET28a(+)中，酶切、测序等鉴定后，重组子pET28a(+)-Sy2转染原核细胞，用载体标签His抗体经SDS-PAGE、Western blot方法检测Sy2重组蛋白的表达。 结果 成功构建出pET28a(+)-Sy2重组质粒，Western blot方法检测出融合蛋白的表达。 结论 成功构建原核表达载体并在原核细胞中表达成包涵体，这种包涵体可以作为抗原制备Sy2的单克隆抗体。

    【关键词】 蛋白磷酸酶;遗传载体;聚合酶链反应

    【Abstract】 Objective To construct and express the recombination vector of pET28a(+)-Sy2(a nov-el Ser/Thr protein phosphatase，named Sy2)plasmid and ready to make Sy2monoclonal antibody for study its biologic function.Methods Sy2phosphatase domain cDNA was amplified by PCR，The frag-ment was digested and recombined into pET28a(+)vector.The prokaryotic vector transfected into BL21(DE3)cell line，and then induced its expression by IPTG.The production of inducing expression was identified by Blotting with anti-tag-his Ab.Results The recombination plasmid was constructed and the fusion protein was expressed successfully.Conclusions pET28a(+)-Sy2plasmid，constructed as a prokaryotic expression vector，could express the Sy2fusion protein.This fusion protein could be used as antigen to development monoclonal antibody against Sy2.It provided a powerful tool for the fu-ture study of biological function of a novel Sy2phosphatase. ......