RESUMO
A series of unnatural base pairs was designed and examined for the expansion of the genetic alphabet and for a better understanding of the mechanism of nucleic acid biosyntheses. To improve the shape complementarity of the previously developed unnatural base pairs, 2-amino-6-(N,N-dimethylamino)purine (x)--pyridon-2-one (y) and 2-amino-6-(2-thienyl)purine (s)--y, the pyrimidine analogue, y, was replaced by a five-member ring, 4-imidazolin-2-one (z), and the s-z pairing in replication was examined. Unnatural bases based on the five-member ring were also applied to the development of non-hydrogen-bonded base pairs.
Assuntos
Código Genético , Oligonucleotídeos/síntese química , Pareamento de Bases , Replicação do DNA , Oligonucleotídeos/química , Nucleotídeos de Purina/química , Nucleotídeos de Pirimidina/químicaRESUMO
Transcription reaction by T7-RNA polymerase was photo-regulated on the basis of two strategies as depicted in Scheme 1. It was found that incorporation reaction of azobenzene-tethered uridine triphosphate proceeded only when azobenzene took trans-form (Scheme 1(A)). On the other hand, the transcription was more efficient when the azobenzene moiety, tethered to the non-template strand of the promoter DNA, was in its cis-form under UV irradiation (Scheme 1(B)). Thus, azobenzene-tethered DNAs are promising for the photo-regulation of gene-expression.
Assuntos
Compostos Azo/química , RNA Polimerases Dirigidas por DNA/metabolismo , DNA/química , Transcrição Gênica/efeitos da radiação , Raios Ultravioleta , Sequência de Bases , Regulação da Expressão Gênica , Modelos Genéticos , Regiões Promotoras Genéticas , Uridina Trifosfato/química , Proteínas ViraisRESUMO
The unnatural base, 2-amino-6-(2-thienyl)purine (designated as s), instead of 2-amino-6-(N,N-dimethylamino)purine (designated as x), was designed in order to improve the specificity and efficiency of the base pairing with pyridin-2-one (designated as y). DNA fragments containing s were chemically synthesized, and the thermal stability and the enzymatic reactions involving the s-y pairing were examined. Thermal denaturation experiments showed that the DNA duplex (12-mer) containing the s-y pair was more stable than that containing the x-y pair. The incorporation of dyTP was also more advantageous to the s-y pairing than the x-y pairing in single-nucleotide insertion experiments using the Klenow fragment of Escherichia coli DNA polymerase I.
