含7-脱氮-2'-脱氧鸟苷的回文寡核苷酸：d[(p)GG*AATTCC]八聚体的固相合成及被核酸内切酶EcoRI识别
Palindromic oligonucleotides containing 7-deaza-2'-deoxyguanosine: solid-phase synthesis of d[(p)GG*AATTCC] octamers and recognition by the endodeoxyribonuclease EcoRI.
作者信息
Seela F, Driller H
出版信息
Nucleic Acids Res. 1986 Mar 11;14(5):2319-32. doi: 10.1093/nar/14.5.2319.
Abstract
Octadeoxynucleotides with the sequence d[(p)GGAATTCC] have been prepared by solid-phase synthesis employing regular and base-modified phosphoramidites. These oligomers which contain an isosterically altered recognition sequence of the endodeoxyribonuclease Eco RI form duplexes under appropriate salt conditions. Since G can represent 7-deaza-2'-deoxyguanosine the oligomers were used as probes to study their cleavage by the endodeoxyribonuclease Eco RI. The enzymatic hydrolysis of the modified octamer was strongly decreased compared to the regular DNA-fragment. This shows that guanine N-7 located at the cleavage site is important for the recognition process by the enzyme. The residual enzymatic activity is discussed on the basis of reduced specificity towards the recognition fragment. The fact that this cleavage occurs already under regular conditions indicates that the process described here bases on an intrinsic property of the oligomer and is different from the star activity.
摘要
通过使用常规和碱基修饰的亚磷酰胺进行固相合成，制备了序列为d[(p)GGAATTCC]的十八脱氧核苷酸。这些含有内切脱氧核糖核酸酶Eco RI等构改变识别序列的寡聚物在适当的盐条件下形成双链体。由于G可以代表7-脱氮-2'-脱氧鸟苷，因此这些寡聚物被用作探针来研究它们被内切脱氧核糖核酸酶Eco RI切割的情况。与常规DNA片段相比，修饰的八聚体的酶促水解作用大大降低。这表明位于切割位点的鸟嘌呤N-7对酶的识别过程很重要。基于对识别片段特异性降低的情况讨论了残余酶活性。这种切割在常规条件下就已经发生，这一事实表明这里描述的过程基于寡聚物的内在特性，并且不同于星号活性。
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