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目的探讨β3GnT8(β3GalT7,AY277592,EC2.4.1.-)基因(人类β3-糖基转移酶家族中的一个新成员)的功能。方法原核表达该蛋白,以纯化的蛋白免疫新西兰大白兔,制备多克隆抗血清;以Westernblot和免疫组化分析其在多种组织细胞的表达情况;并通过荧光免疫细胞化学进行亚细胞定位。结果获得了较纯的表达蛋白和高效价、高特异性的抗血清,并且发现该蛋白大部分位于核膜周围的胞浆中;免疫组化分析显示肿瘤细胞和组织中该酶有较高的表达,与正常组织比较具有一定的差异性。结论该实验获得了β3GnT8/β3GalT7基因的蛋白质和相应抗体,并且对该蛋白进行了亚细胞定位,为对其生物学功能的进一步研究奠定了基础。
Objective To investigate the function of the β3GnT8 (β3GalT7, AY277592, EC2.4.1.-) gene, a new member of the human β3-glycosyltransferase family. Methods The recombinant protein was expressed in prokaryotic cells. The purified protein was used to immunize New Zealand white rabbits to prepare polyclonal antiserum. The expression of polyclonal antiserum was detected by Western blot and immunohistochemistry. The subcellular localization was performed by fluorescence immunocytochemistry. Results The pure expression protein and high titer and high specificity antiserum were obtained, and most of the protein was found in the cytoplasm around the nuclear membrane. Immunohistochemical analysis showed that the enzyme in tumor cells and tissues had higher Expression, compared with normal tissue has a certain difference. Conclusion The protein and corresponding antibody of β3GnT8 / β3GalT7 gene was obtained in this experiment. The subcellular localization of this protein was carried out, which lays a foundation for the further study of its biological function.