EMERGENCE OF A REPLICATING SPECIES FROM AN IN-VITRO RNA EVOLUTION REACTION

被引:69
作者
BREAKER, RR [1 ]
JOYCE, GF [1 ]
机构
[1] Scripps Res Inst, DEPT CHEM, LA JOLLA, CA 92037 USA
关键词
RNA AMPLIFICATION; SELFISH RNA; RNA POLYMERASE; PROMOTER;
D O I
10.1073/pnas.91.13.6093
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
The technique of self-sustained sequence replication allows isothermal amplification of DNA and RNA molecules in vitro. This method relief on the activities of a reverse transcriptase and a DNA-dependent RNA polymerase to amplify specific nucleic acid sequences. We have modified this protocol to allow selective amplification of RNAs that catalyze a particular chemical reaction. During an in vitro RNA evolution experiment employing this modified system, a unique class of ''selfish'' RNAs emerged and replicated to the exclusion of the intended RNAs. Members of this class of selfish molecules, termed RNA Z, amplify efficiently despite their inability to catalyze the target chemical reaction. Their amplification requires the action of both reverse transcriptase and RNA polymerase and involves the synthesis of both DNA and RNA replication intermediates. The proposed amplification mechanism for RNA Z involves the formation of a DNA hairpin that functions as a template for transcription by RNA polymerase, This arrangement links the two strands of the DNA, resulting in the production of RNA transcripts that contain an embedded RNA polymerase promoter sequence.
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页码:6093 / 6097
页数:5
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