Flavell R B, Smith D B
Nucleic Acids Res. 1977 Jul;4(7):2429-44. doi: 10.1093/nar/4.7.2429.
Hyperpolymer formation during the renaturation of DNAs from wheat, calf and E. coli was studied using hydroxyapatite chromatography, electron microscopy and S1 nuclease. Large hyperpolymers could not be eluted from hydroxyapatite with 0.5 M phosphate buffer at 60 degrees C. Large proportions of wheat and E. coli DNAs were incorporated into hyperpolymers when fragments 650 nucleotides long were renatured. A much smaller proportion of calf DNA was incorporated under equivalent conditions. Greater proportions of calf DNA accumulated in hyperpolymers only when longer fragments were incubated. Electron microscopy indicated no obvious differences in the basic structures of hyperpolymers formed by the three DNAs and confirmed the quantitative differences in hyperpolymer formation found by hydroxyapatite chromatography. It is concluded that the proportions and arrangement of the repeated sequences in the chromosomes of higher organisms determine the extent of rapid hyperpolymer formation during DNA renaturation in vitro.
利用羟基磷灰石层析、电子显微镜和S1核酸酶,对来自小麦、小牛和大肠杆菌的DNA复性过程中的超聚合物形成进行了研究。在60℃下,大的超聚合物不能用0.5M磷酸盐缓冲液从羟基磷灰石上洗脱下来。当650个核苷酸长的片段复性时,很大比例的小麦和大肠杆菌DNA被整合到超聚合物中。在相同条件下,只有小得多比例的小牛DNA被整合进去。只有当较长片段孵育时,才有更大比例的小牛DNA积累在超聚合物中。电子显微镜显示,三种DNA形成的超聚合物的基本结构没有明显差异,并证实了羟基磷灰石层析发现的超聚合物形成的数量差异。得出的结论是,高等生物染色体中重复序列的比例和排列决定了体外DNA复性过程中快速超聚合物形成的程度。