Fox K R
Department Physiology & Pharmacology, University of Southampton, UK.
Nucleic Acids Res. 1992 Dec 25;20(24):6487-93. doi: 10.1093/nar/20.24.6487.
The self complementary DNA dodecamers d(CGCGAATTCGCG), d(CGCGTTAACGCG), d(CGCGTATACGCG), d(CGCGATATCGCG), d(CGCAAATTTGCG), d(CGCTTTAAAGCG), d(CGCGGATCCGCG) and d(CGCGGTACCGCG) have been cloned into the Smal site of plasmid pUC19. Radiolabelled polylinker fragments containing these inserts have been digested with nucleases and chemical agents, probing the structure of the central AT base pairs. The sequences AATT and AAATTT are relatively resistant to digestion by DNase I, micrococcal nuclease and hydroxyl radicals, consistent with the suggestion that they possess a narrow minor groove. Nuclease digestion of TTAA is much more even, and comparable to that at mixed sequence DNA. TpA steps in ATAT, TATA and GTAC are cut less well by DNAse I than in TTAA. DNasel cleavage of surrounding bases, especially CpG is strongly influenced by the nature of the central sequence.
自我互补的DNA十二聚体d(CGCGAATTCGCG)、d(CGCGTTAACGCG)、d(CGCGTATACGCG)、d(CGCGATATCGCG)、d(CGCAAATTTGCG)、d(CGCTTTAAAGCG)、d(CGCGGATCCGCG)和d(CGCGGTACCGCG)已被克隆到质粒pUC19的Smal位点。含有这些插入片段的放射性标记多克隆位点片段已用核酸酶和化学试剂消化,以探测中央AT碱基对的结构。序列AATT和AAATTT对DNase I、微球菌核酸酶和羟基自由基的消化相对具有抗性,这与它们具有狭窄小沟的推测一致。TTAA的核酸酶消化更为均匀,与混合序列DNA的消化情况相当。与TTAA相比,DNAse I对ATAT、TATA和GTAC中TpA步的切割效果较差。周围碱基的DNasel切割,尤其是CpG,受到中央序列性质的强烈影响。