Domanskiĭ N N, Galkin A P, Sytnik K M
Ukr Biokhim Zh (1978). 1985 Nov-Dec;57(6):63-8.
A special technique is developed for isolation of biologically functional poly(A)+RNA from higher plant cells to start with some procedures used to isolate RNA of the eucaryote cells. The technique includes cell disruption with a 4 M guanidine thiocyanate solution, hot phenol extraction of RNA following proteinase K digestion and methoxyethanol treatment to remove polysaccharides. Finally, poly(A)+RNA was purified by oligo(dT)-cellulose chromatography. Using this technique poly(A)+RNA preparations were obtained from French bean embryo axis, maize and sugar beet seedings, potato tubers and seedlings. It is shown that poly(A)+RNA isolated from the plant cells directs protein synthesis in the cell-free system. This RNA may be also used as a matrix for synthesis of cDNA in the reverse transcription reaction.
从高等植物细胞中分离具有生物学功能的多聚腺苷酸（poly(A)+）RNA的一种特殊技术，是从一些用于分离真核细胞RNA的程序开始研发的。该技术包括用4M硫氰酸胍溶液破坏细胞，蛋白酶K消化后用热酚提取RNA，并用甲氧基乙醇处理以去除多糖。最后，通过寡聚（dT）-纤维素柱层析纯化多聚腺苷酸（poly(A)+）RNA。利用该技术，从菜豆胚轴、玉米和甜菜幼苗、马铃薯块茎和幼苗中获得了多聚腺苷酸（poly(A)+）RNA制剂。结果表明，从植物细胞中分离的多聚腺苷酸（poly(A)+）RNA可在无细胞系统中指导蛋白质合成。这种RNA也可作为逆转录反应中合成互补DNA（cDNA）的模板。
