- A METHOD FOR THE SITE-SPECIFIC ENZYMATIC LABELLING OF NUCLEIC ACIDS IN VITRO BY INCORPORATION OF UNNATURAL NUCLEOTIDES
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Provided herein are analogs of unnatural nucleotides bearing predominantly hydrophobic nucleobase analogs that form unnatural base pairs during DNA polymerase- mediated replication of DNA or RNA polymerase-mediated transcription of RNA. In this manner, the unnatural nucleobases can be introduced in a site-specific way into oligonucleotides (single or double stranded DNA or RNA), where they can provide for site-specific cleavage, or can provide a reactive linker than can undergo functionalization with a cargo -bearing reagent by means of reaction with a primary amino group or by means of click chemistry with an alkyne group of the unnatural nucleobase linker.
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Paragraph 00176
(2015/02/25)
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- Natural-like replication of an unnatural base pair for the expansion of the genetic alphabet and biotechnology applications
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We synthesized a panel of unnatural base pairs whose pairing depends on hydrophobic and packing forces and identify dTPT3-dNaM, which is PCR amplified with a natural base pair-like efficiency and fidelity. In addition, the dTPT3 scaffold is uniquely tolerant of attaching a propargyl amine linker, resulting in the dTPT3PA-dNaM pair, which is amplified only slightly less well. The identification of dTPT3 represents significant progress toward developing an unnatural base pair for the in vivo expansion of an organism's genetic alphabet and for a variety of in vitro biotechnology applications where it is used to site-specifically label amplified DNA, and it also demonstrates for the first time that hydrophobic and packing forces are sufficient to mediate natural-like replication.
- Li, Lingjun,Degardin, Melissa,Lavergne, Thomas,Malyshev, Denis A.,Dhami, Kirandeep,Ordoukhanian, Phillip,Romesberg, Floyd E.
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p. 826 - 829
(2014/02/14)
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- Furopyridines. XXVII. Reactions of 2-methyl and 2-cyano derivatives of furo[2,3-b]-, -[3,2-b]-, - [2,3-c]- and -[3,2-c]pyridine
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Bromination of 2-methylfuropyridines 1a-d-Me gave the 3-bromo derivatives 2a-d, while the 2-cyano compounds 1a-d-CN resulted in the recovery of the starting compounds. Nitration of 1a-d-Me and 1a-d-CN did not yield the corresponding nitro derivative, except for 1-c-CN giving 3-nitro derivative 3c in 7% yield. N-Oxidation of 1a-d-Me and 1b-d-CN with m- chloroperbenzoic acid yielded the N-oxides 4a-d-Me and 4b-d-CN, whereas 1a- CN did not afford the N-oxide. Cyanation of N-oxides 4a-d-Me and 4b-d-CN with trimethylsilyl cyanide gave the corresponding α-cyanopyridine compounds 5a- d-Me and 5b-d-CN. Chlorination of 4a-d-Me and 4b-d-CN with phosphorus oxychloride also gave the α-chloropyridine compounds 6b-d-Me and 6b-d-CN, accompanying formation of γ-chloropyridine 6a-Me, 6'b-Me and 6'b-CN, β- chloropyridine 6(b)-CN, and α'-chloropyridine derivatives 6'c-Me and 6'c- CN. Acetoxylation of 4a-d-Me and 4b-d-CN with acetic anhydride yielded α- acetoxypyridine compounds 7a-Me and 7b-CN, pyridone compounds 11d-Me, 11c-CN and 11d-CN, 3-acetoxy compounds 8, 9b, 9c, and 2-acetoxymethyl derivatives 10b and 10c.
- Yamaguchi, Seiji,Kurosaki, Masahide,Orito, Keiko,Yokayama, Hajime,Hirai, Yoshiro,Shiotani, Shunsaku
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p. 1237 - 1247
(2007/10/03)
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