Cohen F S, Niles W D
Department of Physiology, Rush Medical College, Chicago, Illinois 61612.
Methods Enzymol. 1993;220:50-68. doi: 10.1016/0076-6879(93)20073-c.
Protocols to reconstitute channels into planar bilayers via fusion methods have now been developed. The greater the intravesicular pressures generated, the greater is the fusion. These pressures can be calculated exactly for any experimental configuration. For some of the configurations, adding nystatin channels to the vesicle membrane will greatly aid fusion. The configurations of the 1990 Method (Figs. 4 and 5) are optimal for fusing vesicles that are reconstituted with ion-selective channels to planar membranes. Greater binding, and ultimately greater fusion, is achieved by ejecting vesicles directly at the membrane rather than by simply adding material to the cis compartment.
目前已经开发出通过融合方法将通道重组到平面双层膜中的方案。产生的囊泡内压力越大，融合程度就越大。对于任何实验配置，这些压力都可以精确计算。对于某些配置，向囊泡膜中添加制霉菌素通道将极大地促进融合。1990年方法的配置（图4和图5）对于将用离子选择性通道重组的囊泡融合到平面膜中是最佳的。通过直接将囊泡喷射到膜上而不是简单地将材料添加到顺式隔室中，可以实现更大的结合，最终实现更大程度的融合。
