Shoyab M, Dastoor M N, Baluda M A
Proc Natl Acad Sci U S A. 1976 May;73(5):1749-53. doi: 10.1073/pnas.73.5.1749.
The integration site of avian myeloblastosis virus (AMV) proviral DNA in DNA from leukemia chicken myeloblasts has been studied by three sequential nucleic acid hybridizations that can localize the proviral DNA according to the repetitiveness of the adjacent cellular DNA regions. First, large denatured cellular DNA fragments (2.1 x 10(6) daltons) were reassociated and fractionated according to sequence reiteration frequenct. Next, DNA remaining single-stranded in each fraction was immobilized on nitrocellulose filters hybridized with an excess of unlabeled 70S RNA from Rous-associated virus-0 to saturate the endogenous proviral DNA sequences.
通过三次连续的核酸杂交研究了禽成髓细胞瘤病毒(AMV)前病毒DNA在白血病鸡成髓细胞DNA中的整合位点,这三次杂交可根据相邻细胞DNA区域的重复情况来定位前病毒DNA。首先,将大的变性细胞DNA片段(2.1×10⁶道尔顿)复性,并根据序列重复频率进行分级分离。接下来,将每个级分中仍为单链的DNA固定在硝酸纤维素滤膜上,与过量的来自劳氏相关病毒0的未标记70S RNA杂交,以使内源性前病毒DNA序列饱和。