کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
2190035 1096233 2006 7 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Revitalization of Six Abandoned Catalytic DNA Species Reveals a Common Three-way Junction Framework and Diverse Catalytic Cores
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی بیولوژی سلول
پیش نمایش صفحه اول مقاله
Revitalization of Six Abandoned Catalytic DNA Species Reveals a Common Three-way Junction Framework and Diverse Catalytic Cores
چکیده انگلیسی

A library containing as many as 1016 nucleic acid candidates is typically used to isolate artificial ribozymes and deoxyribozymes (DNAzymes) in an in vitro selection experiment, with only a handful of sequences surviving many rounds of stringent selection steps. These winning species are generally the focus of interest whereas the less competitive contenders are usually not examined. Nevertheless, molecular species abandoned during the selection process might still represent a rich pool of catalytic motifs that are useful for the examination of DNA's inherent catalytic ability, and for the design of molecular tools for practical applications. Here we report a study of six RNA-cleaving, fluorescence-signaling deoxyribozymes that appeared in the early generations of a previous in vitro selection experiment, using the combined approaches of reselection, rational structural analysis, and reaction condition optimization. All six deoxyribozymes were found to use a three-way junction as a common structural framework for catalysis. However, disparities observed in the conserved nucleotide allocations, methylation interference patterns and metal-ion selectivities, pointed to distinct catalytic cores. The rate constants of the optimized deoxyribozymes fell in the range of ∼0.2 to 1.6 min−1, which are comparable to those of similar ribozymes. Our findings indicate that deoxyribozymes eliminated by harsh selection criteria are structurally simple molecules that can be tailored into efficient catalysts.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Molecular Biology - Volume 357, Issue 3, 31 March 2006, Pages 748–754
نویسندگان
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