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使用纺锤菌素与氯化铯梯度分析法分析DNA及其在多头绒泡菌核糖体DNA限制性核酸酶片段中的应用。

The use of netropsin with CsCl gradients for the analysis of DNA and its application to restriction nuclease fragments of ribosomal DNA from Physarum polycephalum.

作者信息

Matthews H R, Johnson E M, Steer W M, Bradbury E M, Allfrey V G

出版信息

Eur J Biochem. 1978 Jan 16;82(2):569-76. doi: 10.1111/j.1432-1033.1978.tb12052.x.

Abstract

Netropsin binds to DNA in caesium chloride density gradients and reduces the density of the DNA. The DNA is saturated at a netropsin/DNA weight ratio of about 6 and the change in density, deltarho, at saturation is given by deltarho = -109 (dA + dT content)1.87 mg/ml for the six DNAs tested covering dA + dT contents from 0.28 to 0.69. At lower netropsin/DNA ratios the observed density shifts are consistent with a two-site model for netropsin binding to DNA. Netropsin approximately doubles the resolution of Physarum polycephalum nucleolar satellite DNA from main-band DNA. The fragments of P. polycephalum nucleolar satellite DNA obtained with the restriction endonuclease HindIII do not separate on CsCl gradients, even in the presence of netropsin, which shows that the transcribed and non-transcribed sequences in this DNA have similar nucleotide compositions.

摘要

纺锤菌素在氯化铯密度梯度中与DNA结合,并降低DNA的密度。在纺锤菌素/DNA重量比约为6时DNA达到饱和,对于所测试的六种DNA(涵盖dA + dT含量从0.28至0.69),饱和时的密度变化Δρ由Δρ = -109(dA + dT含量)1.87 mg/ml给出。在较低的纺锤菌素/DNA比例下,观察到的密度变化与纺锤菌素结合DNA的双位点模型一致。纺锤菌素使多头绒泡菌核仁卫星DNA与主带DNA的分辨率大约提高了一倍。用限制性内切酶HindIII获得的多头绒泡菌核仁卫星DNA片段在CsCl梯度中无法分离,即使存在纺锤菌素,这表明该DNA中的转录和非转录序列具有相似的核苷酸组成。

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