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高分子量DNA与沉降系数:基于T7噬菌体DNA及两种新型T4噬菌体的新视角

High-molecular-weight DNA and the sedimentation coefficient: a new perspective based on DNA from T7 bacteriophage and two novel forms of T4 bacteriophage.

作者信息

Clark R W, Wever G H, Wiberg J S

出版信息

J Virol. 1980 Jan;33(1):438-48. doi: 10.1128/JVI.33.1.438-448.1980.

Abstract

The DNA molecules from T7 bacteriophage and a recently obtained mutant form of T4D were studied. The DNA of this T4 mutant contains cytosine in place of all of the glucosylated hydroxymethylcytosines normally present in T4. Molecular weights were measured with an electron microscope technique, and sedimentation coefficients were determined in isokinetic sucrose gradients. T7 DNA was found to have an Mr of 26.5 x 10(6). The T4 mutant, which we have termed T4c, produces two distinct phage head and DNA size clases. DNA from the standard heads (T4c DNA) has an Mr of 114.9 x 10(6), and DNA from the petite heads (T4cp DNA) has an Mr of 82.9 x 10(6). This enabled the derivation of an equation of sedimentation coefficient at zero concentration corrected to water at 20 degrees C versus Mr for the molecular weight range of 25 x 10(6) to 115 x 10(6) that is based solely on cytosine-containing DNA standards, thereby avoiding possible anomalies introduced by the glucosylation and hydroxymethylation of cytosine. The theory of Gray et al. provided the best description of the sedimentation coefficient versus Mr relationship, based on the sedimentation coefficients and the molecular weights of the three DNA standards and other evidence.

摘要

对来自T7噬菌体和最近获得的T4D突变体形式的DNA分子进行了研究。这种T4突变体的DNA含有胞嘧啶,取代了T4中通常存在的所有糖基化羟甲基胞嘧啶。用电子显微镜技术测量分子量,并在等速蔗糖梯度中测定沉降系数。发现T7 DNA的Mr为26.5×10⁶。我们将这种T4突变体称为T4c,它产生两种不同的噬菌体头部和DNA大小类别。来自标准头部的DNA(T4c DNA)的Mr为114.9×10⁶,来自小头部的DNA(T4cp DNA)的Mr为82.9×10⁶。这使得能够推导一个仅基于含胞嘧啶的DNA标准品的、在20℃校正至水的零浓度下的沉降系数与Mr的方程,该方程适用于25×10⁶至115×10⁶的分子量范围,从而避免了胞嘧啶糖基化和羟甲基化可能引入的异常。基于三种DNA标准品的沉降系数和分子量以及其他证据,Gray等人的理论对沉降系数与Mr的关系提供了最佳描述。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e0a1/288559/9697e7e2520e/jvirol00169-0461-a.jpg

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