Hybridization of synthetic oligodeoxyribonucleotides to phi chi 174 DNA: the effect of single base pair mismatch.
Hybridization of synthetic oligodeoxyribonucleotides to phi chi 174 DNA: the effect of single base pair mismatch.
复制标题
合成寡脱氧核糖核苷酸与 phi chi 174 DNA 的杂交：单碱基对错配的影响。
DOI：
10.1002/(sici)1096-8628(19960202)61:4
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发表时间：
1979
影响因子：
14.9
通讯作者：
K. Itakura
中科院分区：
文献类型：
--
作者：
R. Wallace;J. Shaffer;R. F. Murphy;J. Bonner;T. Hirose;K. Itakura
通过磷酸三酯法合成了与野生型 (wt) 噬菌体 phi chi 174 的 DNA 互补的寡脱氧核糖核苷酸。寡聚体长度分别为 11、14 和 17 个碱基，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 摄氏度的温度下解离。由于单个错配而引起的热稳定性急剧下降使得可以通过适当选择杂交温度来消除错配双链体的形成。这些结果是关于使用寡核苷酸作为分离特定克隆 DNA 序列的探针进行讨论的。
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.