Catalytic Activity Assay Of The Human UDP-Gal: BetaGlcNAc Beta 1,3-galactosyltransferase Polypeptide 7 Gene And Its Functional Investigation In Tumor Cells

Object: UDP-Gal:betaGlcNAc beta 1,3-galactosyltransferase polypeptide 7 is a new member of glycosyltransferases which belongs to the beta-3 glycosyltransferase family. This thesis aims to the catalytic activity ofβ3GalT7. Simutaneously, study on the relationship between theβ3GalT7 and the tumor cell on the base of the that. Methods: 1.The cDNA gene encodingβ3GalT7, which was amplified from plasmid pGEM-T-T7 by PCR, was subcloned into prokaryotic expression vector pGEX-6p-1, the recombinant plasmid was subsequently transformed into Escherichia coli BL21. Then a fusion protein was expressed after the induction by IPTG. After testified by western blot, we purified the expression products by affinity chromatography. After renaturation, its enzyme activity was assayed by High Performance Liquid Chromatography (HPLC). 2. Theβ3GalT7 fragment was subcloned to the eukaryotic plasmid and transfected to the SGC7901 and A549 cell lines. With fluorescence microscope, laser confocal microscope, RT-PCR, Western blot, the preliminary functions ofβ3GalT7 in the tumor...