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电化学(中英文) ›› 2001, Vol. 7 ›› Issue (1): 25-29. 

• 研究论文 • 上一篇    下一篇

用荧光葡聚糖研究大麦细胞电融合

王明艳,龚叶芳,江志裕   

  1. 连云港市化工高等专科学校化工系,复旦大学生命科学院,复旦大学化学系!上海200433
  • 收稿日期:2001-02-28 修回日期:2001-02-28 出版日期:2001-02-28 发布日期:2001-02-28

Study on Electrofusion of Barley Protoplasts using Fluorescent Dextra

WANG Min yan , GONG Ye fang , JIANG Zhi yu   

  1. (Dept. of Chem., Fudan Univ., Shanghai 200433, China
  • Received:2001-02-28 Revised:2001-02-28 Published:2001-02-28 Online:2001-02-28

摘要: 利用阴离子表面活性物质荧光葡聚糖 (F DX)研究了表面活性物质对大麦细胞电融合的影响 .结果表明 ,F DX可抑制电融合过程 .对放置过大麦细胞原生质体的F DX溶液 ,在荧光显微镜下可观察到其膜表面的荧光圈 ,证明F DX在膜上的吸附 .添加F DX可增加原生质体的电泳速度 ,说明吸附后原生质体表面负电荷增多 .由于相互间静电斥力的增强 ,使细胞的电融合率下降 .此外 ,还利用荧光显微技术研究了细胞电生孔现象 .观察到经电脉冲后溶液中的F DX可进入原生质体内部 ,间接证明了细胞电生孔的存在 .

关键词: 大麦原生质体, 细胞电融合, 电生孔, 膜吸附, 荧光葡聚糖

Abstract: The influence of interfacial active substance on the electrofusion of barley protoplasts was investigated using fluorescent dextra (F DX) as an anion additive. It was found that fluorescent dextra could inhibit the electrofusion process. The adsorption of fluorescent dextra on the membrane of protoplasts was detected by fluorescent microscopy technique. It was observed that after being stored in a solution containing fluorescent dextra a fluorescent ring appeared on the membrane of the protoplast, reflecting the adsorption of fluorescent dextra. The results of electrophoresis experiment also proved the adsorption of F DX on the surface of protoplasts, because the relative mobility of protoplasts increased in the solution containing F DX. The increase of negative charge on the membrane of protoplasts could increase the repulsion between protoplasts, and therefore inhibit the electrofusion process. The electroporation phenomena were also investigated using fluorescent microscopy technique. It was found that F DX could pass into the inside of protoplasts under an electronic pulse. It proves the presence of pores formed in the elecroporation process.

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