Institute of Organic Chemistry and Biochemistry, Czech Academy of Sciences, Flemingovo namesti 2, CZ-16610 Prague 6, Czech Republic.
Org Biomol Chem. 2018 Aug 1;16(30):5427-5432. doi: 10.1039/c8ob01106k.
2'-Deoxyribonucleoside triphosphates (dNTPs) containing 5-(hydroxymethyl)cytosine (5hmC) protected with photocleavable groups (2-nitrobenzyl or 6-nitropiperonyl) were prepared and studied as substrates for the enzymatic synthesis of oligonucleotides and DNA containing a photocaged epigenetic 5hmC base. DNA probes containing photocaged or free 5hmC in the recognition sequence of restriction endonucleases were prepared and used for the study of the photorelease of caged DNA by UV or visible light at different wavelengths. The nitrobenzyl-protected dNTP was a slightly better substrate for DNA polymerases in primer extension or PCR, whereas the nitropiperonyl-protected nucleotide underwent slightly faster photorelease at 400 nm. However, both photocaged building blocks can be used in polymerase synthesis and the photorelease of 5hmC in DNA.
2'-脱氧核苷三磷酸（dNTPs）中含有 5-（羟甲基）胞嘧啶（5hmC），并带有光解保护基团（2-硝基苄基或 6-硝基胡椒基），被用作含有光笼蔽表观遗传 5hmC 碱基的寡核苷酸和 DNA 酶促合成的底物。制备了含有光笼蔽或游离 5hmC 的 DNA 探针，用于在不同波长的紫外光或可见光下研究限制内切酶识别序列中 DNA 的光释放。在引物延伸或 PCR 中，硝基苄基保护的 dNTP 是 DNA 聚合酶稍好的底物，而 6-硝基胡椒基保护的核苷酸在 400nm 处更快地进行光解。然而，这两种光笼蔽砌块都可用于聚合酶合成和 DNA 中 5hmC 的光释放。
