Site-specific biotinylation of RNA molecules by transcription using unnatural base pairs

Moriyama K; Kimoto M; Mitsui T; Yokoyama S; Hirao I

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Site-specific biotinylation of RNA molecules by transcription using unnatural base pairs

机译：RNA分子通过使用非天然碱基对的转录进行位点特异性生物素化

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Direct site-specific biotinylation of RNA molecules was achieved by specific transcription mediated by unnatural base pairs. Unnatural base pairs between 2-amino-6-(2-thienyl)purine (denoted by s) and 2-oxo(1H)pyridine (denoted by y), or 2-amino-6-(2-thiazolyl)purine (denoted as v) and y specifically function in T7 transcription. Using these unnatural base pairs, the substrate of biotinylated-y (Bio-yTP) was selectively incorporated into RNA, opposite s or v in the DNA templates, by T7 RNA polymerase. This method was applied to the immobilization of an RNA aptamer on sensor chips, and the aptamer accurately recognized its target protein. This direct site-specific biotinylation will provide a tool for RNA-based biotechnologies.

机译：RNA分子的直接位点特异性生物素化是通过非天然碱基对介导的特异性转录实现的。 2-氨基-6-（2-噻吩基）嘌呤（以s表示）和2-氧代（1H）吡啶（以y表示）或2-氨基-6-（2-噻唑基）嘌呤（以表示）之间的非天然碱基对如v）和y在T7转录中特异性起作用。使用这些非天然碱基对，通过T7 RNA聚合酶将生物素化y（Bio-yTP）的底物选择性地掺入DNA模板中s或v相对的RNA中。该方法被应用于将RNA适体固定在传感器芯片上，并且该适体准确地识别了其靶蛋白。这种直接的位点特异性生物素化将为基于RNA的生物技术提供工具。

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《Nucleic Acids Research》 |2005年第15期|共8页
作者
Moriyama K; Kimoto M; Mitsui T; Yokoyama S; Hirao I;
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正文语种 eng
中图分类细胞形态学;
关键词
QUARTZ-CRYSTAL MICROBALANCE; DERIVATIVES; COMPONENT; APTAMERS; ACIDS;

机译：石英晶体微平衡;衍生物;组分;适体;酸;

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专利

1. Site-specific biotinylation of RNA molecules by transcription using unnatural base pairs [J] . Moriyama K, Kimoto M, Mitsui T, Nucleic Acids Research . 2005,第15期

机译：RNA分子通过使用非天然碱基对的转录进行位点特异性生物素化
2. Site-specific functionalization of RNA molecules by an unnatural base pair transcription system via click chemistry [J] . Takumi Ishizuka, Michiko Kimoto, Akira Sato, Chemical Communications . 2012,第88期

机译：非自然碱基对转录系统通过点击化学对RNA分子进行位点特异性功能化
3. PCR Amplification and Transcription for Site-Specific Labeling of Large RNA Molecules by a Two-Unnatural-Base-Pair System [J] . MichikoKimoto, RieYamashige, ShigeyukiYokoyama, Journal of Nucleic Acids . 2012,第4期

机译：通过两个非天然碱基对系统对大RNA分子进行位点特异性标记的PCR扩增和转录
4. Development of a Solid-supported Biotinylation Reagent for the Efficient Biotin Labeling to the SH group of Peptides and Small Molecules [C] . Kentarou Fukumoto, Kumi Adachi, Yuri Yamazaki Japanese peptide symposium . 2011

机译：用于高效生物素标记的固体支持的生物素化试剂对SH肽和小分子的高效标记
5. Synthesis of the unnatural amino acids lysine-EDTA and lysine-DTPA, their incorporation into HIV-1 Tat peptides and site-specific mapping on TAR RNA. [D] . Foss, Catherine Anne. 2003

机译：合成非天然氨基酸赖氨酸-EDTA和赖氨酸-DTPA，将其掺入HIV-1 Tat肽中，并在TAR RNA上进行位点特异性定位。
6. Site-specific biotinylation of RNA molecules by transcription using unnatural base pairs [O] . Kei Moriyama, Michiko Kimoto, Tsuneo Mitsui, 2005

机译：RNA分子通过使用非天然碱基对的转录进行位点特异性生物素化
7. Site-specific biotinylation of RNA molecules by transcription using unnatural base pairs [O] . Moriyama Kei, Kimoto Michiko, Mitsui Tsuneo, 2005

机译：RNA分子通过使用非天然碱基对的转录进行位点特异性生物素化

Site-specific biotinylation of RNA molecules by transcription using unnatural base pairs

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