Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, Gilead & IOCB Research Center, Flemingovo nam. 2, CZ-16610, Prague 6, Czech Republic.
Nucleic Acids Res. 2009 Dec;37(22):7612-22. doi: 10.1093/nar/gkp845.
A set of 6 base-modified 2'-deoxyadenosine derivatives was incorporated to diverse DNA sequences by primer extension using Vent (exo-) polymerase and the influence of the modification on cleavage by diverse restriction endonucleases was studied. While 8-substituted (Br or methyl) adenine derivatives were well tolerated by the restriction enzymes and the corresponding sequences were cleaved, the presence of 7-substituted 7-deazaadenine in the recognition sequence resulted in blocking of cleavage by some enzymes depending on the nature and size of the 7-substituent. All sequences with modifications outside of the recognition sequence were perfectly cleaved by all the restriction enzymes. The results are useful both for protection of some sequences from cleavage and for manipulation of functionalized DNA by restriction cleavage.
一套 6 种碱基修饰的 2'-脱氧腺苷衍生物通过引物延伸被 Vent（外切）聚合酶整合到不同的 DNA 序列中，研究了修饰对不同限制内切酶切割的影响。虽然 8 位取代（溴或甲基）腺嘌呤衍生物被限制酶很好地耐受，并且相应的序列被切割，但在识别序列中存在 7 位取代的 7-脱氮腺苷会导致一些酶的切割被阻断，这取决于 7 位取代基的性质和大小。所有修饰位于识别序列之外的序列都被所有限制酶完全切割。这些结果对于保护某些序列免受切割以及通过限制切割来操作功能化 DNA 都非常有用。
