Bioinformatic genome surveys indicate that self-cleaving ribonucleic acids (ribozymes) appear to be widespread among all domains of life, although the functions of only a small number have been validated by biochemical methods. Alternatively, cell-based reporter gene assays can be used to validate ribozyme function. However, reporter activity can be confounded by phenomena unrelated to ribozyme-mediated cleavage of RNA. We established a ribozyme reporter system in Escherichia coli in which a significant reduction of reporter activity is manifest when an active ribozyme sequence is fused to the reporter gene and the expression of a foreign Bacillus subtilis RNaseJ1 5′ exonuclease is induced from a chromosomally-integrated gene in the same cell. The reporter system could be useful for validating ribozyme function in candidate sequences identified from bioinformatics.

中文翻译：

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采用染色体整合的 5' 核酸外切酶基因的基于细胞的核酶报告系统

生物信息学基因组调查表明，自切割核糖核酸（核酶）似乎广泛存在于生命的所有领域，尽管只有少数的功能已通过生化方法得到验证。或者，基于细胞的报告基因检测可用于验证核酶功能。然而，与核酶介导的 RNA 切割无关的现象可能会混淆报告基因的活性。我们在大肠杆菌中建立了一个核酶报告系统，其中当活性核酶序列与报告基因融合并且外源枯草芽孢杆菌 RNaseJ1 5' 核酸外切酶的表达由染色体整合基因诱导时，报告活性显着降低在同一个单元格中。