Laboratoire de Physiologie Végétale, Université de Neuchâtel, Chemin de Chantemerle 20, CH-2000 Neuchâtel, Switzerland.
Plant Physiol. 1984 Jul;75(3):670-4. doi: 10.1104/pp.75.3.670.
A procedure was developed to purify simultaneously peroxisomes and mitochondria from spinach (Spinacia oleracea L.) leaf under isoosmotic and low viscosity conditions. This method involved differential centrifugation and density gradient centrifugation on four layers of Percoll. Chlorophyll-free preparations of highly intact and active organelles were obtained and cross-contamination was negligible. Both organelles were stable for several hours, even if they remained in Percoll. Purified mitochondria were able to carry out the oxidation of different substrates with excellent respiratory control and ADP:O ratios. The method described in the present work was also suitable to purify mitochondria and peroxisomes from potato (Solanum tuberosum L.) tubers.
开发了一种在等渗和低粘度条件下从菠菜(Spinacia oleracea L.)叶中同时纯化过氧化物酶体和线粒体的程序。该方法涉及在四层聚蔗糖上进行差速离心和密度梯度离心。获得了高度完整和活性细胞器的无叶绿素制剂,并且几乎没有交叉污染。即使细胞器留在聚蔗糖中,它们也能稳定数小时。纯化的线粒体能够极好地进行呼吸控制和 ADP:O 比的氧化作用。本工作中描述的方法也适用于从马铃薯(Solanum tuberosum L.)块茎中纯化线粒体和过氧化物酶体。