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Modification of the surface carbohydrate composition of tobacco protoplasts transformed with the human UDP-galactose transporter gene hUGT1

机译:用人UDP半乳糖转运蛋白基因hUGT1转化的烟草原生质体表面碳水化合物组成的修饰

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Tobacco BY-2 cells were transformed with the human UDP-galactose transporter 1 gene (hUGT1) under the control of a 35S promoter. Accumulation of mRNA and protein derived from hUGT1 was detected in the hUGT1-transformed BY-2 cells. To identify the cell surface carbohydrates, BY-2 protoplasts were treated with 21 kinds of biotinylated lectin, stained with fluorescein isothiocyanate (FITC)-conjugated streptavidin and observed by fluorescence microscopy. FITC-ConA and -RCA(120) staining exhibited strong fluorescence on the surface of BY-2 protoplasts, whereas staining with other FITC-lectin conjugates showed faint or no fluorescence. Regarding FITC-ConA and -RCA(120) staining, the fluorescence of hUGT-expressing BY-2 protoplasts was weaker than that of control cells. Decreased FITC fluorescence of hUGT-expressing BY-2 protoplasts was also detected by fluorescence-activated cell sorting (FACS) analysis, suggesting that the surface carbohydrate composition was modified.
机译:在35S启动子的控制下,用人UDP-半乳糖转运蛋白1基因(hUGT1)转化了烟草BY-2细胞。在hUGT1转化的BY-2细胞中检测到了来自hUGT1的mRNA和蛋白质的积累。为了鉴定细胞表面的碳水化合物,用21种生物素化的凝集素处理BY-2原生质体,用异硫氰酸荧光素(FITC)偶联的链霉亲和素染色,并通过荧光显微镜观察。 FITC-ConA和-RCA(120)染色在BY-2原生质体表面显示强荧光,而其他FITC-凝集素缀合物染色则显示微弱或无荧光。关于FITC-ConA和-RCA(120)染色,表达hUGT的BY-2原生质体的荧光比对照细胞弱。还通过荧光激活细胞分选(FACS)分析检测到了表达hUGT的BY-2原生质体FITC荧光的减少,表明表面碳水化合物的组成被修饰。

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