Ruthe H J, Adler J
Biochim Biophys Acta. 1985 Sep 25;819(1):105-13. doi: 10.1016/0005-2736(85)90200-7.
Spheroplasts of Escherichia coli or Salmonella typhimurium were found to fuse in an electric field. We employed the fusion method developed by Zimmermann and Scheurich (1981): Close membrane contact between cells is established by dielectrophoresis (formation of chains of cells by an a.c. field), then membrane fusion is induced by the application of short pulses of direct current. Under optimum conditions the fusion yield was routinely 90%. Fusable spheroplasts were obtained by first growing filamentous bacteria in the presence of cephalexin, then converting these to spheroplasts by the use of lysozyme. The fusion products were viable and regenerated to the regular bacterial form. Fusion of genetically different spheroplasts resulted in strains of bacteria possessing a combination of genetic markers. Fusion could not be achieved with spheroplasts obtained by growing the cells in the presence of penicillin or by using lysozyme on bacteria of usual size.
人们发现大肠杆菌或鼠伤寒沙门氏菌的原生质球在电场中会发生融合。我们采用了齐默尔曼和朔伊里希（1981年）开发的融合方法：通过介电泳（利用交流电场形成细胞链）使细胞间建立紧密的膜接触，然后施加短脉冲直流电诱导膜融合。在最佳条件下，融合率通常为90%。可融合的原生质球是通过先在头孢氨苄存在的情况下培养丝状细菌，然后使用溶菌酶将其转化为原生质球而获得的。融合产物具有活力，并能再生为正常的细菌形态。遗传上不同的原生质球融合产生了具有多种遗传标记组合的细菌菌株。用在青霉素存在的情况下培养细胞所获得的原生质球，或者对正常大小的细菌使用溶菌酶所获得的原生质球，无法实现融合。
