Ranjekar P K, Pallotta D, Lafontaine J G
Biochim Biophys Acta. 1976 Feb 18;425(1):30-40. doi: 10.1016/0005-2787(76)90213-6.
Barley and wheat DNAs have been characterized by studying their kinetics of reassociation, melting properties and sedimentation behaviour in neutral CsCl gradients as well as in Cs2SO4 gradients containing Ag+ or Hg2+. In both species, reassociation kinetics have revealed the presence of approx. 76% redundant nucleotide sequences which have been grouped into very rapidly reassociating (Cot 0-0.01), rapidly reassociating (Cot 0.01-1.0) and slowly reassociating (Cot 1-100) fractions. The barley Cot 0-0.01 and Cot 0.01-1.0 fractions as well as the wheat Cot 0.01-1.0 fraction form narrow bands upon centrifugation in CsCl gradients. Under similar experimental conditions both Cot 0.01 and Cot 1.0-100 wheat fractions and the barley Cot 1.0-100 fraction form broad bands each having several shoulders. Thermal denaturation studies of most of the above reassociated fractions have shown a considerable degree of order in their duplexes with an average hyperchromicity of 21.5%. When native, high molecular weight barley DNA is centrifuged in Ag+/CS2SO4 density gradients (RF = 0.2), two satellites appear on the heavier side of the main band, as against one in the case of wheat. The two minor peaks, designated as satellites I and II, have buoyant densities of 1.702 and 1.698 g/cm3, respectively, in neutral CsCl gradients and together represent about 8-9% of total barley DNA. Upon centrifugation in Hg2+/CS2SO4 density gradients, one satellite is observed in both barley and wheat and it accounts for 1-2% of their genomes.
通过研究大麦和小麦DNA在中性CsCl梯度以及含有Ag⁺或Hg²⁺的Cs₂SO₄梯度中的重缔合动力学、解链特性和沉降行为,对它们进行了表征。在这两个物种中,重缔合动力学表明存在约76%的冗余核苷酸序列,这些序列已被分为重缔合非常快(Cot 0 - 0.01)、重缔合快(Cot 0.01 - 1.0)和重缔合慢(Cot 1 - 100)的组分。大麦的Cot 0 - 0.01和Cot 0.01 - 1.0组分以及小麦的Cot 0.01 - 1.0组分在CsCl梯度中离心时形成窄带。在相似的实验条件下,小麦的Cot 0.01和Cot 1.0 - 100组分以及大麦的Cot 1.0 - 100组分均形成宽带,每个宽带都有几个肩峰。对上述大多数重缔合组分的热变性研究表明,它们的双链体具有相当程度的有序性,平均增色效应为21.5%。当天然的高分子量大麦DNA在Ag⁺/CS₂SO₄密度梯度(RF = 0.2)中离心时,在主带较重的一侧出现两条卫星带,而小麦只有一条。这两个小峰分别称为卫星I和卫星II,在中性CsCl梯度中的浮力密度分别为1.702和1.698 g/cm³,它们总共约占大麦总DNA的8 - 9%。在Hg²⁺/CS₂SO₄密度梯度中离心时,大麦和小麦均观察到一条卫星带,它占其基因组的1 - 2%。