Uckert W, Fleischhacker M, Kettmann R
Virology. 1986 Dec;155(2):742-6. doi: 10.1016/0042-6822(86)90236-9.
Infection of a human lymphoblastoid B cell line (Raji cells) with type D retroviruses, originally isolated either from subhuman primates (MPMV, LV) or from permanent human cell lines (PMFV, HeLaV, HEp-2V) led to the production of type D retrovirus particles. Subsequent cocultivation of uninfected and virus-producing Raji cells was employed for the generation of sufficient amounts of covalently closed circular DNA molecules (cccDNA). Highest amounts of cccDNA were obtained after cocultivation of virus-producing Raji cells and homologous uninfected cells at a ratio of 1 to 3 for 72 hr. The cccDNAs of type D retroviruses migrated at about 4.3 kbp compared to lambda DNA/HindIII markers. Digestion of cccDNAs with restriction endonucleases which have one recognition site generated molecules of approximately 8 kbp. The restriction endonuclease site analysis of the cccDNA of type D retroviruses revealed a genomic heterogeneity among the different isolates.
用最初从非人灵长类动物（猴泡沫病毒、淋巴细胞脉络丛脑膜炎病毒）或人类永久细胞系（人泡沫病毒、HeLa病毒、HEp - 2病毒）分离出的D型逆转录病毒感染人淋巴母细胞样B细胞系（拉吉细胞），会导致产生D型逆转录病毒颗粒。随后，将未感染和产生病毒的拉吉细胞进行共培养，以产生足够数量的共价闭合环状DNA分子（cccDNA）。在将产生病毒的拉吉细胞与同源未感染细胞以1比3的比例共培养72小时后，获得了最高量的cccDNA。与λDNA/HindIII标记相比，D型逆转录病毒的cccDNA在约4.3千碱基对处迁移。用具有一个识别位点的限制性内切酶消化cccDNA会产生约8千碱基对的分子。D型逆转录病毒cccDNA的限制性内切酶位点分析揭示了不同分离株之间的基因组异质性。
