In vitro selection of self-cleaving RNAs with a low pH optimum

被引:60
作者
Jayasena, VK [1 ]
Gold, L [1 ]
机构
[1] NEXSTAR PHARMACEUT INC, BOULDER, CO 80301 USA
关键词
D O I
10.1073/pnas.94.20.10612
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
RNAs that undergo a sapid site-specific cleavage at low pH have been selected by in vitro selection (the SELEX process), The cleavage does not require the addition of any divalent metal ions, and is in fact inhibited by divalent metal ions, spermine, or high concentrations of monovalent metal ions, This low pH catalyzed cleavage results in a 2',3'-cyclic phosphate at the 3' end and a free hydroxyl at the 5' end, The reaction proceeds with a calculated rate of 1.1 min(-1) at room temperature in cacodylate buffer at pH 5.0. The rate of cleavage is dependent on the pH and shows an optimum around pH 4.0. The rate constant is independent of RNA concentration, indicating to an intramolecular reaction, Autocatalytic cleavage at low pH, in the absence of a metal ion requirement, adds to the reaction possibilities that may have existed on the prebiotic earth.
引用
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页码:10612 / 10617
页数:6
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