Hélène C
Laboratoire de Biophysique, INSERM U 201, CNRS UA 481, Muséum National d'Histoire Naturelle, Paris, France.
Curr Opin Biotechnol. 1993 Feb;4(1):29-36. doi: 10.1016/0958-1669(93)90028-u.
Single sites within long double-helical DNA molecules can be recognized by a variety of mechanisms. Different strategies have been used to adapt sequence-specific recognition to sequence-specific cleavage of duplex DNA. Any nucleic acid can be converted into an artificial nuclease by the attachment of a cleaving reagent. Alternatively, a sequence-specific ligand can be used to protect a methylase recognition site from methylation. The protected site may then be cleaved selectively by a restriction endonuclease (the so-called 'Achilles heel' cleavage technique). Recent developments in this area have shown that it is possible to cleave chromosomal DNA at single sites within bacterial and eukaryotic genomes.
长双链DNA分子内的单个位点可通过多种机制识别。人们已采用不同策略使序列特异性识别适应双链DNA的序列特异性切割。通过连接切割试剂,任何核酸都可转化为人工核酸酶。另外,可使用序列特异性配体保护甲基化酶识别位点不被甲基化。然后,受保护的位点可被限制性内切酶选择性切割(即所谓的“阿喀琉斯之踵”切割技术)。该领域的最新进展表明,有可能在细菌和真核生物基因组内的单个位点切割染色体DNA。