Shaw J E, Levinger L F, Carter C W
J Virol. 1979 Feb;29(2):657-65. doi: 10.1128/JVI.29.2.657-665.1979.
Micrococcal nuclease digestion was used to analyze Epstein-Barr virus (EBV) DNA structure in nuclei of transformed cells. Digests of virus-producing (P3HR-1), non-virus-producing (Raji), and superinfected Rajii cell nuclei were fractionated by electrophoresis on agarose gels, transferred to nitrocellulose, and hybridized to 32P-labeled EBV DNA. The viral DNA of Raji nuclei produced a series of bands on electrophoresis whose lengths were integral multiples of a unit size, which was the same as the repeat length of host DNA. Viral DNA in nuclei of P3HR-1 and superinfected Raji cells produced faintly visible bands superimposed on a smear of viral DNA which dominated the hybridization pattern. No differences were detected in the patterns when total DNA digests from Raji, P3HR-1, and an EBV DNA-negative cell line (U-698M) were analyzed by ethidium bromide staining or by hybridization with the use of 32P-labeled lymphoblastoid cell DNA as probe. We conclude that the EBV episomal DNA of Raji cells is folded into nucleosomes, whereas most of the viral DNA of P3HR-1 and superinfected Raji cells is not. This pattern of DNA organization differs signficantly from that in papova group viruses.
使用微球菌核酸酶消化法分析转化细胞细胞核中爱泼斯坦-巴尔病毒(EBV)的DNA结构。对产生病毒的(P3HR-1)、不产生病毒的(Raji)以及超感染的Raji细胞的细胞核消化产物,通过琼脂糖凝胶电泳进行分级分离,转移至硝酸纤维素膜上,并与32P标记的EBV DNA杂交。Raji细胞核的病毒DNA在电泳时产生一系列条带,其长度是一个单位大小的整数倍,这与宿主DNA的重复长度相同。P3HR-1细胞核和超感染的Raji细胞中的病毒DNA产生的条带隐约可见,叠加在主导杂交图谱的病毒DNA涂片上。当用溴化乙锭染色或使用32P标记的淋巴母细胞DNA作为探针进行杂交分析Raji、P3HR-1和EBV DNA阴性细胞系(U-698M)的总DNA消化产物时,未检测到模式上的差异。我们得出结论,Raji细胞的EBV附加体DNA折叠成核小体,而P3HR-1和超感染的Raji细胞的大多数病毒DNA则没有。这种DNA组织模式与乳头瘤病毒组病毒的模式有显著差异。