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A cell-based ribozyme reporter system employing a chromosomally-integrated 5 ‘ exonuclease gene
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Document Title
A cell-based ribozyme reporter system employing a chromosomally-integrated 5 ' exonuclease gene
Author
Aroonsri A, Kongsee J, Gunawan JD, Aubry DA, Shaw PJ
Name from Authors Collection
Affiliations
National Science & Technology Development Agency - Thailand; National Center Genetic Engineering & Biotechnology (BIOTEC); Indonesia International Institute for Life-Sciences (i3L)
Type
Article
Source Title
BMC MOLECULAR AND CELL BIOLOGY
Year
2021
Volume
22
Open Access
gold, Green Published
Publisher
BMC
DOI
10.1186/s12860-021-00357-7
Format
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.
Keyword
coli cell-based system | E | glmS riboswitch | Hammer-head ribozyme | Reporter system | Ribozyme | RNaseJ1
Industrial Classification
Knowledge Taxonomy Level 1
Knowledge Taxonomy Level 2
Knowledge Taxonomy Level 3
Funding Sponsor
Thailand Research Fund (TRF); National Science and Technology Development Agency (NSTDA) [TRG608001, P17-51076]
License
CC BY
Rights
Authors
Publication Source
WOS