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爱泼斯坦-巴尔病毒的DNA。II. 爱泼斯坦-巴尔病毒株DNA限制性内切酶片段的分子量比较及B95-8株末端片段的鉴定

DNA of Epstein-Barr virus. II. Comparison of the molecular weights of restriction endonuclease fragments of the DNA of Epstein-Barr virus strains and identification of end fragments of the B95-8 strain.

作者信息

Hayward S D, Kieff E

出版信息

J Virol. 1977 Aug;23(2):421-9. doi: 10.1128/JVI.23.2.421-429.1977.

Abstract

Incubation of the DNA of the B95-8 strain of Epstein-Barr virus [EBV (B95-8) DNA] with EcoRI, Hsu I, Sal I, or Kpn I restriction endonuclease yielded 8 to 15 fragments separable on 0.4% agarose gels and ranging in molecular weight from less than 1 to more than 30 x 10(6). Bam I and Bgl II yielded fragments smaller than 11 x 10(6). Preincubation of EBV (B95-8) DNA with lambda exonuclease resulted in a decrease in the Hsu I A and Sal I A and D fragments, indicating that these fragments are positioned near termini. The electrophoretic profiles of the fragments produced by cleavage of the DNA of the B95-8, HR-1, and Jijoye strains of EBV were each distinctive. The molecular weights of some EcoRI, Hsu I, and Sal I fragments from the DNA of the HR-1 strain of EBV [EBV (HR-1) DNA] and of EcoRI fragments of the DNA of the Jijoye strain of EBV were identical to that of fragments produced by cleavage of EBV (B95-8) DNA with the same enzyme, whereas others were unique to each strain. Some Hsu I, EcoRI, and Sal I fragments of EBV (HR-1) DNA and Kpn I fragments of EBV (B95-8) DNA were present in half-molar abundance relative to the majority of the fragments. In these instances, the sum of the molecular weights of the fragments was in excess of 10(8), the known molecular weight of EBV (HR-1) and (B95-8) DNA. The simplest interpretation of this finding is that each EBV (HR-1), and possibly also (B95-8), DNA preparation contains two populations of DNA molecules that differ in the arrangement of DNA sequences about a single point, such as has been described for herpes simplex virus DNA. Minor fragments could also be observed if there were more than one difference in primary structure of the DNAs. The data do not exclude more extensive heterogeneity in primary structure of the DNA of the HR-1 strain. However, the observation that the relative molar abundance of major and minor fragments of EBV (HR-1) DNA did not vary between preparations from cultures that had been maintained separately for several years favors the former hypothesis over the latter.

摘要

将爱泼斯坦 - 巴尔病毒B95 - 8株的DNA [EBV (B95 - 8) DNA] 与EcoRI、Hsu I、Sal I或Kpn I限制性内切酶一起温育,产生了8至15个片段,这些片段在0.4%琼脂糖凝胶上可分离,分子量范围从小于1×10⁶至大于30×10⁶。Bam I和Bgl II产生的片段小于11×10⁶。用λ外切核酸酶对EBV (B95 - 8) DNA进行预温育导致Hsu I A以及Sal I A和D片段减少,这表明这些片段位于末端附近。由EBV的B95 - 8、HR - 1和Jijoye株的DNA切割产生的片段的电泳图谱各有特点。来自EBV的HR - 1株的DNA [EBV (HR - 1) DNA] 的一些EcoRI、Hsu I和Sal I片段以及EBV的Jijoye株的DNA的EcoRI片段的分子量与用相同酶切割EBV (B95 - 8) DNA产生的片段的分子量相同,而其他片段则是每个株系所特有的。EBV (HR - 1) DNA的一些Hsu I、EcoRI和Sal I片段以及EBV (B95 - 8) DNA的Kpn I片段相对于大多数片段以半摩尔丰度存在。在这些情况下,片段的分子量总和超过了10⁸,即EBV (HR - 1) 和 (B95 - 8) DNA的已知分子量。对这一发现最简单的解释是,每个EBV (HR - 1),可能还有 (B95 - 8) 的DNA制剂都包含两种DNA分子群体,它们在围绕单个点的DNA序列排列上有所不同,就像单纯疱疹病毒DNA所描述的那样。如果DNA的一级结构存在不止一个差异,也可能观察到小片段。这些数据并不排除HR - 1株DNA一级结构存在更广泛的异质性。然而,来自分别保存数年的培养物的制剂中,EBV (HR - 1) DNA的主要和小片段的相对摩尔丰度没有变化这一观察结果更支持前一种假设而非后一种。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5e5a/515844/4412c7d0113f/jvirol00212-0223-a.jpg

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