绿色荧光蛋白基因导入胡萝卜愈伤组织并获得表达

目的:将绿色荧光蛋白基因(green fluorescent protein,GFP)重组到胡萝卜愈伤组织细胞中,使其获得表达,为今后利用GFP基因作为植物报告基因提供条件。方法:通过冻融法将含有GFP基因的重组表达载体PBI1121转入到根癌农杆菌EHA105中,再利用根癌农杆菌介导的方法将GFP基因导入到胡萝卜愈伤组织细胞中,经过除菌和抗性筛选后观测转化结果。结果:荧光显微镜观测到被转化的愈伤组织在受蓝光激发后发出绿色荧光,利用PCR法扩增出约740bp的目的基因片断。结论:GFP基因在胡萝卜愈伤组织细胞中获得了表达。

Transformation of Green Fluorescent Protein Gene into the Carrot Tissue and Expression of the Gene

Objectives: To recombine the green fluorescent protein(GFP) into the tissue of the carrot and obtain the expresser,the aim was to find the condition for the GFP gene as the plant report gene.Methods: The combined express vector PBI121 which contained the GFP gene,was transformed into the Agribacterium tumefaciens EHA105,and the GFP gene was introduced into the tissue of carrot by Agribacterium tumefaciens,observing the results after removing the EHA105 and giving the fastness filter.Results: By the observation of fluorescent microscope using the blue light can the transformed tissue emit the green fluorescence,and using the PCR analysis a section of 740bp was observed.Conclusion: The GFP gene was proved expression in the tissue of carrot.