Aiyada Aroonsri1, Jindaporn Kongsee2, Jeremy David Gunawan3, Daniel Abidin Aubry3, Philip James Shaw2. 1. National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Pathum Thani, 12120, Thailand. aiyada.aro@biotec.or.th. 2. National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Pathum Thani, 12120, Thailand. 3. School of Life Science, Indonesia International Institute for Life Sciences (i3L), Jakarta, 13210, Indonesia.
Abstract
BACKGROUND: 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. RESULTS: 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. CONCLUSIONS: The reporter system could be useful for validating ribozyme function in candidate sequences identified from bioinformatics.
BACKGROUND: 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. RESULTS: 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. CONCLUSIONS: The reporter system could be useful for validating ribozyme function in candidate sequences identified from bioinformatics.
Authors: S Fujita; T Koguma; J Ohkawa; K Mori; T Kohda; H Kise; S Nishikawa; M Iwakura; K Taira Journal: Proc Natl Acad Sci U S A Date: 1997-01-21 Impact factor: 11.205