Shoyab M, Baluda M A
J Virol. 1975 Oct;16(4):783-9. doi: 10.1128/JVI.16.4.783-789.1975.
The distribution of oncornavirus DNA sequences in various tissues of normal chickens and of chickens with leukemia or kidney tumors induced by avian myeloblastosis virus (AMV) was analyzed by DNA-RNA hybridization using 35S AMV RNA as a probe. All the tissues from normal chickens which were tested contained the same average cellular concentration of endogenous oncornavirus DNA. In contrast, different tissues from lekemic chickens and from chickens bearing kidney tumors contained different concentrations of AMV homologous DNA: in some tissues there was no increase whereas other tissues acquired additional AMV-specific DNA sequences. The increase was the greatest in tissues which can become neoplastic after infection, such as myeloblasts, erythrocytes, and kidney cells. It was directly demonstrated that DNA from AMV-induced kidney tumor contains AMV sequences which are absent in DNA from normal cells. A similar finding had been previously obtained with leukemic cells (15). 3H-labeled 35S RNA from purified AMV was exhaustively hybridized with an excess of normal chicken DNA to remove all the viral RNA sequences which are complementary to DNA from uninfected cells. The 3H-labeled RNA which failed to hybridize was isolated by hydroxylapatite column chromatography which separates DNA-RNA hybrids from single-stranded RNA. The residual RNA hybridized to chicken kidney tumor DNA but did not rehybridize with normal chicken DNA.
以35S禽成髓细胞瘤病毒(AMV)RNA为探针,通过DNA - RNA杂交分析了正常鸡以及由AMV诱发白血病或肾肿瘤的鸡的各种组织中肿瘤病毒DNA序列的分布情况。所有接受检测的正常鸡组织中,内源性肿瘤病毒DNA的平均细胞浓度相同。相比之下,患白血病的鸡和患肾肿瘤的鸡的不同组织中,AMV同源DNA的浓度不同:一些组织中没有增加,而其他组织获得了额外的AMV特异性DNA序列。在感染后可能发生肿瘤的组织中,如成髓细胞、红细胞和肾细胞,增加最为明显。直接证明了AMV诱发的肾肿瘤的DNA中含有正常细胞DNA中不存在的AMV序列。此前在白血病细胞中也得到了类似的发现(15)。将纯化的AMV的3H标记的35S RNA与过量的正常鸡DNA进行彻底杂交,以去除所有与未感染细胞的DNA互补的病毒RNA序列。未杂交的3H标记RNA通过羟基磷灰石柱色谱法分离,该方法可将DNA - RNA杂交体与单链RNA分开。残留的RNA与鸡肾肿瘤DNA杂交,但不与正常鸡DNA重新杂交。