Naumov G N
Mol Gen Mikrobiol Virusol. 1985 Sep(9):44-7.
Chromatographic methods have been used to purify the DNA of plasmid RP1. DNA was purified in two stages. DNA was precipitated by ethanol and separated from RNA and proteins in Sepharose 4B column after lysis of plasmid containing cells by alkaline solution of sodium dodecylsulphate. Separation of the total DNA preparation and isolation of plasmid DNA was achieved at the second stage by chromatography on the hydroxyapatite column. The resulting purified plasmid DNA was free of RNA, protein and linear fragments of chromosomal DNA. The plasmid DNA kept intact native structure and possessed the transforming activity. The DNA of RP1 yield after purification by the described technique presented 70-80 micrograms per g of wet biomass.
色谱法已被用于纯化质粒RP1的DNA。DNA分两个阶段进行纯化。通过乙醇沉淀DNA，并在含有质粒的细胞被十二烷基硫酸钠碱性溶液裂解后，在琼脂糖4B柱上与RNA和蛋白质分离。在第二阶段，通过在羟基磷灰石柱上进行色谱法实现总DNA制剂的分离和质粒DNA的分离。所得纯化的质粒DNA不含RNA、蛋白质和染色体DNA的线性片段。质粒DNA保持完整的天然结构并具有转化活性。通过所述技术纯化后，RP1的DNA产量为每克湿生物质70 - 80微克。
