Yamaguchi J, Lim P Y, Aratani K, Akazawa T
Research Institute for Biochemical Regulation, School of Agricultural Sciences, Nagoya University, Japan.
Cell Struct Funct. 1992 Apr;17(2):87-92. doi: 10.1247/csf.17.87.
A method has been developed to isolate pure preparations of nuclei in high yield from commercially available viable rice embryos (germ), employing extraction with buffer solution containing glycerol (without detergent) and polyamine, followed by centrifugation on a 30% Percoll cushion. The intactness of the isolated nuclei was confirmed by light microscopy as well as electron microscopy. The protein profiles of both whole nuclei and nuclear extracts obtained by SDS-PAGE, organellar marker enzyme activities, DNA and RNA analyses, and in vitro RNA synthesis, all indicate that the highly purified nuclei are isolated from rice embryos.
已开发出一种方法，可从市售的活水稻胚（胚芽）中高产率地分离出纯净的细胞核制剂，该方法采用含甘油（无去污剂）和多胺的缓冲溶液进行提取，然后在30%的Percoll垫层上进行离心。通过光学显微镜和电子显微镜确认了分离出的细胞核的完整性。通过SDS-PAGE获得的全细胞核和核提取物的蛋白质谱、细胞器标记酶活性、DNA和RNA分析以及体外RNA合成，均表明从水稻胚中分离出了高度纯化的细胞核。
