Wallace R B, Shaffer J, Murphy R F, Bonner J, Hirose T, Itakura K
Nucleic Acids Res. 1979 Aug 10;6(11):3543-57. doi: 10.1093/nar/6.11.3543.
Oligodeoxyribonucleotides complementary to the DNA of the wild type (wt) bacteriophage phi chi 174 have been synthesized by the phosphotriester method. The oligomers, 11, 14, and 17 bases long, are complementary to the region of the DNA which accounts for the am-3 point mutation. When hybridized to am-3 DNA, the oligonucleotides form duplexes with a single base pair mismatch. The thermal stability of the duplexes formed between wt and am-3 DNAs has been measured. The am-3 DNA:oligomer duplexes dissociate at a temperature about 10 degrees C lower than the corresponding wt DNA:oligomer duplexes. This dramatic decrease in thermal stability due to a single mismatch makes it possible to eliminate the formation of the mismatched duplexes by the appropriate choice of hybridization temperature. These results are discussed with respect to the use of oligonucleotides as probes for the isolation of specific cloned DNA sequences.
通过磷酸三酯法合成了与野生型（wt）噬菌体φX174的DNA互补的寡脱氧核糖核苷酸。这些长度为11、14和17个碱基的寡聚物与导致am-3点突变的DNA区域互补。当与am-3 DNA杂交时，寡核苷酸形成具有单个碱基对错配的双链体。已测量了wt和am-3 DNA之间形成的双链体的热稳定性。am-3 DNA：寡聚物双链体在比相应的wt DNA：寡聚物双链体低约10℃的温度下解链。由于单个错配导致的热稳定性的显著降低使得通过适当选择杂交温度来消除错配双链体的形成成为可能。针对使用寡核苷酸作为探针分离特定克隆DNA序列的情况讨论了这些结果。
