科研之友
微信
新浪微博
Facebook
分享链接
摘要
In an integrated cell system, mitochondria play a pivotal role in energy metabolism and the maintenance of cellular homeostasis. In most cases, protocols for the purification of the mitochondrial fraction are based on blender-homogenisation of the selected tissue and differential centrifugation of the cell homogenate in a Percoll (TM) gradient. The method of homogenisation usually represents the critical step which affects the yield and quality of the mitochondria isolated from plant cells. To isolate mitochondria from minute, mg-amounts of plant tissue (e.g., pollen tubes), we have developed a new method for the homogenisation of pollen tubes using a 30-mu m pore size cell strainer. The isolated mitochondria were shown to be pure, intact, and functional, based on marker enzyme assays, transmission electron microscopy, and flow cytometry analysis, respectively. Furthermore, the isolated mitochondria were used to extract mitochondrial DNA and to analyse mitochondrial proteins. The new method was easy, reliable, and efficient.
- 出版日期2011-7
- 单位南京农业大学
