Zajac-Kaye M, Brown E, Friend C
Virus Res. 1986 Oct;6(1):45-55. doi: 10.1016/0168-1702(86)90056-0.
The integration pattern, copy number and DNA expression of the Friend virus genome was examined in Friend erythroleukemia cells induced to differentiate with dimethyl sulfoxide, hexamethylene bisacetamide and sodium butyrate. The integrated proviral DNA in Friend erythroleukemia cells was examined by Southern hybridization with a cloned Friend virus (F-MuLV) probe at days 1 and 4 following inducer treatment, as well as on day 6 at which time the cells had remained for 48 h in inducer free medium. KpnI fragments 9 and 5.7 kb long were observed. The copy number of each fragment remained constant throughout the erythroid differentiation process. EcoRI digestion of DNA isolated from cells at different times following the inducer treatment demonstrated multiple integration sites of the proviral genome, which also remained constant during the differentiation process. Proviral DNA expression was examined at 4 h and 4 days following inducer treatment as well as on day 6 by which time cells remained for 48 h in inducer free medium. Northern blot hybridization to the F-MuLV probe indicated no change in the provirus gene expression independent of the class of inducers. These observations reinforce our conclusions that the viral genome and its transcription product do not play a major role in the differentiation process of Friend erythroleukemic cells.
在经二甲基亚砜、六亚甲基双乙酰胺和丁酸钠诱导分化的弗氏红白血病细胞中,检测了弗氏病毒基因组的整合模式、拷贝数和DNA表达。在诱导剂处理后的第1天和第4天,以及在第6天(此时细胞已在无诱导剂培养基中培养48小时),通过用克隆的弗氏病毒(F-MuLV)探针进行Southern杂交,检测弗氏红白血病细胞中整合的前病毒DNA。观察到长度为9 kb和5.7 kb的KpnI片段。在整个红系分化过程中,每个片段的拷贝数保持恒定。对诱导剂处理后不同时间从细胞中分离的DNA进行EcoRI酶切,结果表明前病毒基因组存在多个整合位点,这些位点在分化过程中也保持不变。在诱导剂处理后的4小时和4天,以及在第6天(此时细胞已在无诱导剂培养基中培养48小时)检测前病毒DNA表达。用F-MuLV探针进行Northern印迹杂交表明,无论诱导剂类别如何,前病毒基因表达均无变化。这些观察结果强化了我们的结论,即病毒基因组及其转录产物在弗氏红白血病细胞的分化过程中不发挥主要作用。