BMC Molecular and Cell Biology | |
A cell-based ribozyme reporter system employing a chromosomally-integrated 5′ exonuclease gene | |
article | |
Aroonsri, Aiyada1  Kongsee, Jindaporn1  Gunawan, Jeremy David2  Aubry, Daniel Abidin2  Shaw, Philip James1  | |
[1] National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA);School of Life Science, Indonesia International Institute for Life Sciences (i3L) | |
关键词: Ribozyme; E; coli cell-based system; glmS riboswitch; Hammer-head ribozyme; RNaseJ1; Reporter system; | |
DOI : 10.1186/s12860-021-00357-7 | |
学科分类:内科医学 | |
来源: Colegio Oficial de Psicologos | |
【 摘 要 】
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.
【 授权许可】
CC BY|CC0
【 预 览 】
Files | Size | Format | View |
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RO202108090002667ZK.pdf | 1492KB | download |