Fluorescence-based high-throughput assay for human DNA (cytosine-5)-methyltransferase 1.
Fluorescence-based high-throughput assay for human DNA (cytosine-5)-methyltransferase 1.
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DOI：
10.1016/j.ab.2010.02.032
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发表时间：
2010-06-01
影响因子：
2.9
通讯作者：
Stivers JT
中科院分区：
文献类型：
--
作者：
Ye Y;Stivers JT
We have developed the first economical and rapid nonradioactive assay method that is suitable for high-throughput screening of the important pharmacological target human DNA (cytosine-5)-methyltransferase I (DNMT1). The method combines three key innovations: the use of a truncated form of the enzyme that is highly active on a 26 base pair hemimethylated DNA duplex substrate, the introduction of the methylation site into the recognition sequence of a restriction endonuclease, and the use of a fluorogenic readout method. The extent of DNMT1 methylation is reflected in the protection of the DNA substrate from endonuclease cleavage, which would otherwise result in a large fluorescence increase. The assay has been validated in a high-throughput format and trivial changes in the substrate sequence and endonuclease allow adaptation of the method to any bacterial or human DNA methyltransferase.
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影响因子：
5.6
作者：
Handa, V;Jeltsch, A
通讯作者：
Jeltsch, A
影响因子：
4.8
作者：
Bacolla, A;Pradhan, S;Wells, RD
通讯作者：
Wells, RD
影响因子：
4.3
作者：
Galili, Naomi;Cerny, Jan;Raza, Azra
通讯作者：
Raza, Azra
DOI：
10.1002/tera.1420500504
发表时间：
1994-11-01
期刊：
TERATOLOGY
影响因子：
--
作者：
ROGERS, JM;FRANCIS, BM;CHERNOFF, N
通讯作者：
CHERNOFF, N
DOI：
10.1073/pnas.91.25.11797
发表时间：
1994-12-06
影响因子：
11.1
作者：
JUTTERMANN, R;LI, E;JAENISCH, R
通讯作者：
JAENISCH, R