Waksman Institute of Microbiology, Rutgers University Piscataway, New Jersey 08854.
Plant Physiol. 1980 Aug;66(2):291-4. doi: 10.1104/pp.66.2.291.
Chloroplasts can be obtained by gentle lysis or mild shear of spheroplasts of vitamin B(12)-deficient Euglena gracilis and then purified by isopycnic sedimentation on gradients of Ludox AM or Percoll. The chloroplasts appear compact and highly refractile by phase contrast or Hoffmann contrast microscopy. Upon incubation with [(3)H]leucine or [(35)S]methionine, the chloroplasts incorporate the amino acids into protein at rates that are 100-fold faster than we had previously observed with Euglena and up to 8-fold faster than with chloroplasts of spinach. Euglena chloroplasts prepared by the current procedure are thus qualitatively superior to those previously available from Euglena and at least as active in protein synthesis as chloroplasts from higher plants.
叶绿体可以通过温和裂解或轻微剪切维生素 B(12)缺乏的衣藻球形体来获得,然后通过在 Ludox AM 或 Percoll 梯度上的等密度沉降来纯化。叶绿体在相差或霍夫曼对比显微镜下呈现出紧凑和高度折射的外观。在用 [(3)H]亮氨酸或 [(35)S]甲硫氨酸孵育时,叶绿体将氨基酸掺入蛋白质的速度比我们以前用衣藻观察到的速度快 100 倍,比菠菜叶绿体快 8 倍。因此,通过当前程序制备的衣藻叶绿体在质量上优于以前从衣藻获得的叶绿体,并且在蛋白质合成方面至少与高等植物的叶绿体一样活跃。