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牛乳腺炎DNA的剖析。I. 四种分子群体的限制性内切酶图谱,这些分子群体在其长链和短链成分的相对方向上有所不同。

Anatomy of bovine mammillitis DNA. I Restriction endonuclease maps of four populations of molecules that differ in the relative orientation of their long and short components.

作者信息

Buchman T G, Roizman B

出版信息

J Virol. 1978 Jan;25(1):395-407. doi: 10.1128/JVI.25.1.395-407.1978.

Abstract

In this paper, we report that the DNA of bovine mammillitis virus (BMV) consists of two covalently linked components that are 71.5 x 10(6) and 15.7 x 10(6) in molecular weight and designated L and S, respectively. We further report that the BMV DNA consists of four equimolar populations differing only in the orientation of the L and S components relative to each other. This conclusion is based on the following: (i) The sum molecular weight of fragments generated by digestion of BMV DNA with Hsu I, Hpa I, Bgl II, or Xba I significantly exceeds the established molecular weight of the intact DNA. (ii) In each digest, the fragments form three groups differing in molar concentration. In reference to the molar concentration of intact DNA, each enzyme digest contained a set of four fragments 0.25 M in concentration, a set of four fragments 0.5 M in concentration, and a variable size set, unique for each enzyme digest, 1.0 M in concentration. (iii) Experiments involving digestion of intact DNA by lambda exonuclease followed by restriction endonuclease digestion established that each of four 0.5 M fragments were positioned at the termini of the BMV DNA. (iv) Complete maps for the fragments generated by each enzyme established that the 0.25 M fragments arise by fusion of the sequences of the terminal fragments when these are juxtaposed as a consequence of the inversion of L and S components. The maps also established the dimensions of the L and S components. We conclude that the structure of BMV DNA is similar to that of HSV DNA previously shown to consist of two unequal size components that invert relative to each other.

摘要

在本文中,我们报告牛乳头炎病毒(BMV)的DNA由两个共价连接的组分组成,其分子量分别为71.5×10⁶和15.7×10⁶,分别命名为L和S。我们还报告BMV DNA由四个等摩尔群体组成,它们之间的区别仅在于L和S组分彼此相对的方向。这一结论基于以下几点:(i)用Hsu I、Hpa I、Bgl II或Xba I消化BMV DNA产生的片段的总分子量明显超过完整DNA已确定的分子量。(ii)在每个消化产物中,片段形成三组,摩尔浓度不同。参照完整DNA的摩尔浓度,每种酶消化产物都包含一组浓度为0.25M的四个片段、一组浓度为0.5M的四个片段以及一组大小可变的片段(每种酶消化产物独特),浓度为1.0M。(iii)涉及用λ外切核酸酶消化完整DNA然后进行限制性内切核酸酶消化的实验确定,四个0.5M片段中的每一个都位于BMV DNA的末端。(iv)每种酶产生的片段的完整图谱确定,0.25M片段是由末端片段序列融合产生的,当L和S组分倒置导致这些末端片段并列时。这些图谱还确定了L和S组分的尺寸。我们得出结论,BMV DNA的结构与先前显示由两个大小不等的组分组成且彼此相对倒置的单纯疱疹病毒(HSV)DNA的结构相似。

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