Patch C T, Lewis A M, Levine A S
Proc Natl Acad Sci U S A. 1972 Nov;69(11):3375-9. doi: 10.1073/pnas.69.11.3375.
The complementary DNA strands of the nondefective adenovirus 2 (Ad2)-simian virus 40 (SV40) hybrid virus, Ad2(+)ND(1), were separated by isopycnic banding in a CsCl density gradient in the presence of synthetic polyribonucleotides. Separated strands were used in DNA-RNA hybridization reactions with RNA from cells productively infected by Ad2 or SV40, and with complementary SV40 RNA transcribed asymmetrically in vitro. About five times as much Ad2 RNA hybridized to the light stand of Ad2(+)ND(1) as to the heavy strand. Complementary RNA and early SV40 RNA (RNA synthesized before viral DNA replication) had significant homology only with the light strand. Only half as much of a preparation of RNA synthesized before and after viral DNA replication (early-plus-late SV40 RNA) hybridized to the light strand as to the heavy strand. These results indicate that templates for both late and early SV40 RNA are present in Ad2(+)ND(1). Therefore, the small SV40 segment within this virus (10-18% of the SV40 genome) must contain a transcription-control region. Ad2(+)ND(1) should thus be useful in the selective study of transcription as it occurs in cells infected by the oncogenic virus SV40.
在合成多聚核糖核苷酸存在的情况下，通过氯化铯密度梯度中的等密度离心法，分离出无缺陷腺病毒2（Ad2）-猴病毒40（SV40）杂交病毒Ad2(+)ND(1)的互补DNA链。将分离出的链用于DNA-RNA杂交反应，分别与被Ad2或SV40有效感染的细胞中的RNA，以及体外不对称转录的互补SV40 RNA进行杂交。与Ad2(+)ND(1)轻链杂交的Ad2 RNA量大约是与重链杂交量的五倍。互补RNA和早期SV40 RNA（病毒DNA复制前合成的RNA）仅与轻链有显著同源性。病毒DNA复制前后合成的RNA制剂（早期加晚期SV40 RNA）与轻链杂交的量只有与重链杂交量的一半。这些结果表明，Ad2(+)ND(1)中存在晚期和早期SV40 RNA的模板。因此，该病毒内的小SV40片段（占SV40基因组的10-18%）必须包含一个转录控制区。因此，Ad2(+)ND(1)在对致癌病毒SV40感染的细胞中发生的转录进行选择性研究中应该是有用的。
