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Constructing an efficient trans-acting genomic HDV ribozyme
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DOI：10.1016/0014-5793(96)00941-6.png)
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摘要
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We have engineered a genomic HDV ribozyme to construct several tr ans-acting ribozymes for use in trans to cleave target RNAs. Among the 10 different combinations attempted, only HDV88-Trans had cleavage activity on the 13-nucleotide substrate, R13, in vitro, To improve the cleavage efficiency, at least in vitro, of the HDV88-Trans ribozyme (k(clv)=0.022 min(-1)), we have constructed several variants that differ in forming stem II (length) in the pseudoknot secondary structure model, When cleavage rate constants were analyzed and compared among variants of HDV88-Trans, HDV88-Trans-4 yielded k(clv)=1.7 min(-1), HDV88-Trans-4 thus represents the highest active genomic HDV ribozyme that functions in trans thus far constructed, and has activity under physiological conditions (pH 7.1 at 37 degrees C with 1 mM of MgCl2).
Keyword:
HDV ribozyme
pseudoknot structure
ribozyme, trans-acting
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IF:
3
论文数:
2.3W
被引数:
3.8W
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