Isolation of episomal bovine papillomavirus chromatin and identification of a DNase I-hypersensitive region
Isolation of episomal bovine papillomavirus chromatin and identification of a DNase I-hypersensitive region
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游离型牛乳头瘤病毒染色质的分离和 DNase I 超敏感区域的鉴定
DOI：
10.1128/jvi.46.2.567-574.1983
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发表时间：
1983
影响因子：
5.4
通讯作者：
G. Sauer
中科院分区：
文献类型：
--
作者：
F. Rösl;W. Waldeck;G. Sauer
乳头瘤病毒染色质的研究一直受到阻碍的组织培养系统，这些病毒的营养生长的可用性。因此，我们使用了牛乳头瘤病毒1型转化的仓鼠胚胎成纤维细胞，每个细胞含有200至250个附加体基因组当量作为病毒染色质的来源。显示选择性分离的染色质比在蔗糖密度梯度中共沉淀的成熟猿猴病毒40染色质稍大（80S）。Fo I和Fo II均存在于牛乳头瘤病毒1型染色质中。一个快速沉降的部分，其结构仍然是未知的，也包含寡聚牛乳头瘤病毒1型DNA。通过原位DNase消化分离的细胞核和随后的切割的牛乳头瘤病毒1型DNA与各种限制性内切酶，一个主要的DNase超敏感区域中检测到的染色质。该区域包含约320个碱基对，位于相对物理图谱位置0.88和0.92之间。
The investigation of papillomavirus chromatin has been hampered by the unavailability of a tissue culture system for vegetative growth of these viruses. We have used, therefore, bovine papillomavirus type 1-transformed hamster embryo fibroblasts containing 200 to 250 episomal genome equivalents per cell as a source of viral chromatin. The selectively isolated chromatin was shown to be slightly larger (80S) than the mature simian virus 40 chromatin, which was cosedimented in a sucrose density gradient. Both Fo I and Fo II were present in the bovine papillomavirus type 1 chromatin. A fast-sedimenting fraction, whose structure is still unknown, also contained oligomeric bovine papillomavirus type 1 DNA. By in situ DNase digestion of isolated nuclei and subsequent cleavage of the bovine papillomavirus type 1 DNA with various restriction endonucleases, a major DNase-hypersensitive region was detected in the chromatin. This region, comprising approximately 320 base pairs, is located between the relative physical map positions 0.88 and 0.92.