ANATOMY OF HERPES-SIMPLEX VIRUS DNA - STRAIN DIFFERENCES AND HETEROGENEITY IN LOCATIONS OF RESTRICTION ENDONUCLEASE CLEAVAGE SITES
ANATOMY OF HERPES-SIMPLEX VIRUS DNA - STRAIN DIFFERENCES AND HETEROGENEITY IN LOCATIONS OF RESTRICTION ENDONUCLEASE CLEAVAGE SITES
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DOI：
10.1073/pnas.72.5.1768
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发表时间：
1975-01-01
影响因子：
11.1
通讯作者：
ROIZMAN, B
中科院分区：
文献类型：
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作者：
HAYWARD, GS;FRENKEL, N;ROIZMAN, B
用HinIII或Eco RI限制性内切酶消化单纯疱疹病毒DNA，得到11至15个分子量在1 × 10(6)至28 × 10(6)之间的片段。在0.3%琼脂糖凝胶中，单纯疱疹病毒1型的DNA片段与4种单纯疱疹病毒2型的DNA片段的电泳图谱可以很容易地区分开来；然而，对于每种血清型，实验室菌株之间以及在人类宿主外传代最少次数的分离株之间存在显著差异。用任何一种酶消化单纯疱疹病毒的所有dna都可产生两类摩尔浓度不同的片段（主要和次要）。此外，虽然一个完整的单纯疱疹1(F1) DNA分子的分子量约为98 × 10(6)，但所有主要和次要的HinIII片段的分子量总和为160 × 10(6)，仅7个主要片段的分子量仅为60 × 10(6)。这些不寻常的特征表明，在单个分子的解理位点位置存在有限的异质性。我们已经排除了小片段是由于病毒连续未稀释传代时出现的高变异密度缺陷DNA污染引起的可能性。事实上，后一种类型的DNA对HinIII的切割具有抗性，只产生了两种EcoRI片段；这表明有缺陷的分子是由病毒DNA一小段的许多串联重复序列组成的。正常密度的病毒DNA的异质性似乎与分子的结构组织有关，并不一定意味着遗传含量的差异。
Digestion of herpes simplex virus DNA by the HinIII or Eco RI restriction endonucleases yielded 11 to 15 fragments with molecular weights between 1 x 10(6) and 28 x10(6). The electrophoretic profiles obtained in 0.3% agarose gels with DNA fragments from none different strains of herpes simplex virus type 1 could be readily differentiated from the patterns exhibited by the corresponding fragments from four separate strains of type 2 virus; however, with each serotype, the laboratory strains differed significantly among themselves and also from isolates passaged a minimum number of times outside the human host. Digestion of all DNAs of herpes simples virus with either enzyme reproducibly generated two classes of fragments (major and minor) which differed in molar ocncentration. Moreover, although the molecular weight of an intact herpes simplex 1(F1) DNA molecule is approximately 98 x 10(6), the summed molecular weights of all major and minor HinIII fragments totalled 160 x 10(6), and the seven major fragments alone accounted for only 60 x 10(6). These unusual features indicate the existence of limited heterogeneity in the positions of cleavage sitet along individual molecules. We have eliminated the possibility that minor fragments arose from contamination with the defective DNA of high byoyant density which appears on serial undiluted passage of the virus. In fact, this latter type of DNA was resistant to cleavage by HinIII and gave large amounts of only two species of EcoRI fragments; suggesting that the defective molecules consist of many tandem repeats of a small segment of viral DNA. The heterogeneity in the viral DNA of normal density appears to be related to the structural organization of the molecules and does not necessarily imply differences in genetic content.