Protein synthesis by isolated plant mitochondria.

摘要: Publisher Summary This chapter describes the procedures to isolate essentially sterile plant mitochondria that synthesize protein in an energy-dependent, chloramphenicol-sensitive but cycloheximide-insensitive process. The spectrum of proteins synthesized by isolated is same as those labeled vivo presence cycloheximide. Mitochondria capable synthesis vitro have been from a range tissues including fleshy roots and tubers, etiolated seedling tissue, green tissue. major prerequisite are uncontaminated with other subcellular structures or bacteria morphologically functionally intact. Purification carried out centrifugation on either sucrose gradients silica-sol gradient material, Percoll. Sucrose routinely used for purification nongreen they considerably more effective than Percoll removal contaminating micro-bodies. Maximal rates dependent isolation intact, coupled free bacterial contamination. High specific activity [35S]methionine substrate mitochondrial synthesis.