Nayak D P
Proc Natl Acad Sci U S A. 1974 Apr;71(4):1164-8. doi: 10.1073/pnas.71.4.1164.
The kinetics of hybrid formation between the RNA of BrdU-activated endogenous guinea pig virus and the DNA of leukemic, normal, or BrdU-activated guinea pig cells were measured by the technique of RNA.DNA hybridization in DNA excess. The results suggest that virus-specific sequences representing some 60-70% of the viral genome are unique (2-3 copies per haploid cell genome), while the remainder (30-40%) are reiterated (147 copies), and that the reiterated virus-specific DNA may be composed of more than one species, each having a different reiteration frequency. No difference was found in the quantity of viral DNA sequences contained in normal, leukemic, or bromodeoxyuridine-activated guinea pig cells. These data are considerably different from those reported for exogenous (infectious) oncornaviruses, where cells infected or transformed by exogenous RNA tumor viruses have been shown to contain increased amounts of virus-specific DNA. The data reported here are consistent with the contention that preexisting viral genes are activated by bromodeoxyuridine treatment. Results of hybridization experiments done at different DNA/RNA ratios suggest that although the virus-specific DNA is partly unique and partly reiterated, the viral RNA does not contain any detectable internal reiteration. Total mass of the viral RNA sequences is around 0.7 to 1 x 10(7) daltons.
采用DNA过量时的RNA-DNA杂交技术,测定了BrdU激活的内源性豚鼠病毒RNA与白血病、正常或BrdU激活的豚鼠细胞DNA之间杂交形成的动力学。结果表明,代表病毒基因组约60%-70%的病毒特异性序列是独特的(单倍体细胞基因组中每个有2-3个拷贝),而其余部分(30%-40%)是重复的(147个拷贝),并且重复的病毒特异性DNA可能由不止一种组成,每种具有不同的重复频率。在正常、白血病或溴脱氧尿苷激活的豚鼠细胞中所含病毒DNA序列的数量未发现差异。这些数据与报道的外源性(感染性)肿瘤病毒的数据有很大不同,在外源性RNA肿瘤病毒感染或转化的细胞中,已显示含有增加量的病毒特异性DNA。这里报道的数据与预先存在的病毒基因被溴脱氧尿苷处理激活的观点一致。在不同DNA/RNA比率下进行的杂交实验结果表明,尽管病毒特异性DNA部分是独特的,部分是重复的,但病毒RNA不包含任何可检测到的内部重复。病毒RNA序列的总质量约为0.7至1×10⁷道尔顿。