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植物基因组分析。II. 大麦(Hordeum vulgare)和小麦(Triticum aestivum)中重复DNA的特征分析。

Analysis of the genome of plants. II. Characterization of repetitive DNA in barley (Hordeum vulgare) and wheat (Triticum aestivum).

作者信息

Ranjekar P K, Pallotta D, Lafontaine J G

出版信息

Biochim Biophys Acta. 1976 Feb 18;425(1):30-40. doi: 10.1016/0005-2787(76)90213-6.

Abstract

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.

摘要

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%。

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