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三种低DNA含量禾本科植物中的重复DNA

Repetitive DNA in three Gramineae species with low DNA content.

作者信息

Deshpande V G, Ranjekar P K

出版信息

Hoppe Seylers Z Physiol Chem. 1980 Aug;361(8):1223-33. doi: 10.1515/bchm2.1980.361.2.1223.

Abstract

The genomes of three Gramineae species, namely finger millet (Eleusine coracana), pearl millet (Pennisetum americanum) and rice (Oryza sativa) are characterized by studying their DNA denaturation-reassociation properties. The reassociation kinetics measurement of the sonicated DNA (500--700 nucleotide pairs) indicate the presence of a heterogeneous, repetitive DNA fraction accounting for 49--54% of the total DNA in all three species. From the cot 1/2 value of the slow reassociating DNA, the genome size is estimated as 3.0 X 10(8) np in finger millet, 7.8 X 10(8) np in pearl millet and 9.0 X 10(8) np in rice. The melting patterns of the total DNAs reveal Tm value of 88.6 degrees C in the case of pearl millet and 85.0 degrees C in the case of finger millet and rice. Total repetitive and cot 1.0 DNA fractions in all the three species are isolated and their melting properties are compared with those of respective sonicated DNAs. In finger millet, the Tm values of cot 25 and cot 1 fractions are lower by 10.8 degrees C and 12.8 degrees C, respectively, than that of sonicated DNA and thus exhibit the presence of a base pair mismatch in the range of 10.8--12.8%. In rice, the Tm values of the fractions cot 50 and cot 1 are slightly lower than that of sonicated DNA and reveal a nucleotide mismatching of only 1.8--3.8%. In the case of pearl millet cot 10 DNA fraction a high-melting DNA component (Tm = 92 degrees C) representing 12% of the total cot 10 DNA and a low-melting component with a Tm of 78 degrees C are present. In cot 1 DNA fraction of pearl millet the proportion of the high-melting component is 35% and it has a Tm or 94.8 degrees C. Optical reassociation studies of cot 1.0 DNA fractions have revealed the presence of two kinetically distinct components, namely minor fast-reassociating and major slow-reassociating, having complexities in the range of 330--390 np and 1.28 X 10(5)--6.0 X 10(5) np, respectively in pearl millet and rice and only one DNA fraction with an average complexity of 7.7 X 10(4) np in finger millet.

摘要

通过研究三种禾本科植物,即龙爪稷(Eleusine coracana)、珍珠粟(Pennisetum americanum)和水稻(Oryza sativa)的DNA变性-复性特性,对它们的基因组进行了表征。对超声处理的DNA(500-700个核苷酸对)进行的复性动力学测量表明,在所有这三个物种中都存在异质的、重复的DNA组分,占总DNA的49%-54%。根据慢复性DNA的初始浓度一半时的杂交时间(cot 1/2)值,估计龙爪稷的基因组大小为3.0×10⁸个核苷酸对,珍珠粟为7.8×10⁸个核苷酸对,水稻为9.0×10⁸个核苷酸对。总DNA的解链模式显示珍珠粟的解链温度(Tm)值为88.6℃,龙爪稷和水稻的Tm值为85.0℃。分离出所有三个物种的总重复DNA和初始浓度为1.0时的DNA组分,并将它们的解链特性与各自超声处理的DNA的解链特性进行比较。在龙爪稷中,初始浓度为25时的DNA组分和初始浓度为1时的DNA组分的Tm值分别比超声处理的DNA低10.8℃和12.8℃,因此显示出碱基对错配的比例在10.8%-12.8%之间。在水稻中,初始浓度为50时的DNA组分和初始浓度为1时的DNA组分的Tm值略低于超声处理的DNA,显示出核苷酸错配仅为1.8%-3.8%。在珍珠粟初始浓度为10时的DNA组分中,存在一个高熔点DNA组分(Tm = 92℃),占总初始浓度为10时的DNA的12%,以及一个Tm为78℃的低熔点组分。在珍珠粟初始浓度为1时的DNA组分中,高熔点组分的比例为35%,其Tm为94.8℃。对初始浓度为1.0时的DNA组分进行的光学复性研究表明,存在两个动力学上不同的组分,即较小的快速复性组分和较大的慢速复性组分,在珍珠粟和水稻中其复杂度分别在330-390个核苷酸对和1.28×10⁵-6.0×10⁵个核苷酸对范围内,而在龙爪稷中只有一个平均复杂度为7.7×10⁴个核苷酸对的DNA组分。

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