β-1,4-半乳糖基转移酶Ⅱ、Ⅴ表达的亚细胞结构定位研究

为了研究β-1,4-半乳糖基转移酶 和 (β-1,4-Gal T- and )蛋白表达的亚细胞结构定位 ,本实验构建了β-1,4-Gal T- 和 融合绿色荧光蛋白 (GFP)表达质粒 ,分别将构建的质粒转染到 PC12细胞和肝癌 772 1细胞中 ,在荧光显微镜下观察β-1,4-Gal T- 和 在其中表达的亚细胞结构定位。发现 ,β-1,4-Gal T- 和 主要表达在这两种细胞的细胞核旁的 Golgi复合体 ,说明它们主要分布在 Golgi复合体上。提示它们可能是在 Golgi复合体参与蛋白质的糖链修饰

SUBCELLULAR LOCALIZATIONS OF β-1,4-GALACTOSYLTRANSFERASE- Ⅱ AND Ⅴ

In order to study the subcellular localizations of β-1,4-galactosyltransferase Ⅱ and Ⅴ(β-1,4-GalT-Ⅱ and Ⅴ), the enhanced green fluorescent protein (EGFP) segment was attached to the carboxyl terminus of human β-1,4-GalT-Ⅱ and Ⅴ by standard recombinant techniques. The cDNA fragments of the full-length β-1,4-GalT-Ⅱ and Ⅴ were generated by polymerse chain reaction and then cloned into pEGFPN3 vector to create the plasmids of pEGFPN3-β-1,4-GalT-Ⅱ and pEGFPN3-β-1,4-GalT-Ⅴ. After the two plasmids were transfected into PC12 and hepatocellular carcinoma cells SMMC-7721, the subcellular localizations of β-1,4-GalT-Ⅱ and Ⅴ were analyzed in terms of the fusion green fluorescent protein detected by fluorescent microscopy. β-1,4-GalT-Ⅱ and Ⅴ were mainly located in the Golgi complex. The results suggested that β-1,4-GalT-Ⅱ and Ⅴ might take part in modifying the sugar chains of glycoproteins in the Golgi complex.