N4-acyl-2'-deoxycytidine-5'-triphosphates for the enzymatic synthesis of modified DNA.
N4-acyl-2'-deoxycytidine-5'-triphosphates for the enzymatic synthesis of modified DNA.
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DOI：
10.1093/nar/gky435
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发表时间：
2018-07-06
影响因子：
14.9
通讯作者：
Meškys R
中科院分区：
文献类型：
--
作者：
Jakubovska J;Tauraite D;Birštonas L;Meškys R
各种各样的修饰核碱基被用作研究DNA和RNA的工具。由于实际原因，最适合修饰的位置是嘧啶的C5和嘌呤的C7。不幸的是，通过仅使用这两个位置，不能将修饰的核苷酸的库扩展到最大。在这里，我们展示了新的N4-酰化2′-脱氧胞苷核苷酸（dCAcyl）的合成和酶促掺入。我们发现多种家族A和B DNA聚合酶有效地使用dCAcylTP作为底物。当Taq、Klenow片段（exo-）、Bsm和KOD XL DNA聚合酶用于引物延伸反应时，除了形成互补的CAcyl·G对之外，N4-酰基胞嘧啶和腺嘌呤之间还发生强碱基配对。相比之下，校正phi 29 DNA聚合酶成功地利用dCAcylTP，但在相同条件下易于形成CAcyl·A碱基对。此外，我们表明，末端脱氧核苷酸转移酶是能够纳入多达几百个N4-酰化脱氧胞苷核苷酸。这些数据揭示了新的N4-酰化脱氧胞苷核苷酸作为修饰的DNA的酶促合成的有益底物，其可以进一步应用于DNA片段的特异性标记、适体的选择或光固定。
A huge diversity of modified nucleobases is used as a tool for studying DNA and RNA. Due to practical reasons, the most suitable positions for modifications are C5 of pyrimidines and C7 of purines. Unfortunately, by using these two positions only, one cannot expand a repertoire of modified nucleotides to a maximum. Here, we demonstrate the synthesis and enzymatic incorporation of novel N4-acylated 2′-deoxycytidine nucleotides (dCAcyl). We find that a variety of family A and B DNA polymerases efficiently use dCAcylTPs as substrates. In addition to the formation of complementary CAcyl•G pair, a strong base-pairing between N4-acyl-cytosine and adenine takes place when Taq, Klenow fragment (exo–), Bsm and KOD XL DNA polymerases are used for the primer extension reactions. In contrast, a proofreading phi29 DNA polymerase successfully utilizes dCAcylTPs but is prone to form CAcyl•A base pair under the same conditions. Moreover, we show that terminal deoxynucleotidyl transferase is able to incorporate as many as several hundred N4-acylated-deoxycytidine nucleotides. These data reveal novel N4-acylated deoxycytidine nucleotides as beneficial substrates for the enzymatic synthesis of modified DNA, which can be further applied for specific labelling of DNA fragments, selection of aptamers or photoimmobilization.
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DOI：
10.1016/0027-5107(90)90162-w
发表时间：
1990-11-01
期刊：
MUTATION RESEARCH
影响因子：
--
作者：
BASU, AK;ESSIGMANN, JM
通讯作者：
ESSIGMANN, JM
影响因子：
3.2
作者：
Bohacova, Sona;Ludvikova, Lucie;Hocek, Michal
通讯作者：
Hocek, Michal
影响因子：
11.4
作者：
Delarue, M;Boulé, JB;Papanicolaou, C
通讯作者：
Papanicolaou, C
影响因子：
64.8
作者：
HUNTER, WN;BROWN, T;KENNARD, O
通讯作者：
KENNARD, O
影响因子：
16.6
作者：
Hollenstein, Marcel;Hipolito, Christopher;Perrin, David M.
通讯作者：
Perrin, David M.