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Volumn 286, Issue 5437, 1999, Pages 123-126

Imidazole rescue of a cytosine mutation in a self-cleaving ribozyme

Author keywords

[No Author keywords available]

Indexed keywords

ADENINE; CYTOSINE; IMIDAZOLE; RIBONUCLEOPROTEIN; RIBOZYME;

EID: 0033215448     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.286.5437.123     Document Type: Article
Times cited : (263)

References (31)
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    • a of the base and that, in the precursor, the N-3 of γC75 may be positioned to accept the proton from the 2′-hydroxyl group of the ribose at position -1.
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    • 32P]triphosphate and polynucleotide kinase after dephosphorylating with calf intestinal phosphatase (6). The mutations at position 76 and modifications to the sequence 5′ to the cleavage site were generated by oligonucleotide- directed mutagenesis on single-stranded uracil-containing templates of the plasmid (6).
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    • note
    • -1). Buffers that were tested and failed to support activity were tris-HCl, Hepes, Pipes, and MES at pH 7.5; CHES at pH 8.5; and sodium acetate at a variety of pH values. Other potential bases - ethylenediamine (pH 7.8), pyridine (pH 7.5), and ammonia (ammonium acetate, pH 8.5) - failed to rescue cleavage activity. The tris buffer did not inhibit imidazole-dependent cleavage of the C76u ribozyme. However, significant stimulation by imidazole was not a general phenomenon for the HDV antigenomic ribozyme. With the wild-type ribozyme, a moderate increase in the rate constant (1.2-fold) occurred with the addition of 200 mM imidazole (pH 7.4) to the reaction mixture (Table 1). We observed no stimulation of cleavage activity with the C76a ribozyme.
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    • note
    • Supported by NIH grant GM47322. We thank T. Cech, S. Crary, C. Fierke, D. Herschlag, T.-s. Hsieh, and T. Wadkins for critical comments and helpful suggestions.


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.