论文部分内容阅读
目的:探讨P9-ZFD基因片段在人骨骼肌中表达的蛋白质。方法:以MG骨骼肌RNA为模板,扩增编码P9-ZFD片段的cDNA,构建pFT24a-P9-ZFD表达载体,经BL21(DE3)诱导表达P9-ZFD蛋白和组氨酸亲和层析法进行纯化,并制备P9-ZFD抗体。Western blot鉴定MG和对照组骨骼肌中与P9-ZFD抗体产生特异性免疫反应的蛋白组分。结果:骨骼肌中与P9-ZFD抗体产生特异性免疫反应的蛋白质相对分子质量约40000,在伴胸腺增生或胸腺瘤MG骨骼肌中的表达水平明显高于对照组(P<0.001)。结论:骨骼肌中40 000的蛋白是P9-ZFD基因片段的编码产物,该蛋白在伴胸腺增生或胸腺瘤MG骨骼肌中的表达水平明显上调,可能具有重要的病理生理意义。
Objective: To investigate the expression of P9-ZFD gene in human skeletal muscle. Methods: cDNA of P9-ZFD fragment was amplified by using MG skeletal muscle RNA as a template, and pFT24a-P9-ZFD expression vector was constructed. P9-ZFD protein was induced by BL21 (DE3) and histidine affinity chromatography Purified, and P9-ZFD antibody was prepared. Western blot was used to identify the protein components that specifically immunoreacted with P9-ZFD antibody in skeletal muscle of MG and control groups. Results: The relative molecular mass of protein with specific immune response to P9-ZFD in skeletal muscle was about 40 000, which was significantly higher in skeletal muscle with or without thymoma than in control group (P <0.001). CONCLUSION: The 40 000 protein in skeletal muscle is the coding product of P9-ZFD gene. The protein is significantly up-regulated in thymus hyperplasia or MG skeletal muscle and may have important pathophysiological significance.