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A cell-based ribozyme reporter system employing a chromosomally-integrated 5' exonuclease gene

BMC Mol Cell Biol. 2021-03; 
Aiyada Aroonsri, Jindaporn Kongsee, Jeremy David Gunawan, Daniel Abidin Aubry, Philip James Shaw
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Gene Synthesis a synthetic DNA containing the Bacillus subtilis rnjA gene sequence (NCBI Gene ID: 939483) with in-frame C-terminal flexible linker (GGSGGGSGG) and a six histidine residue tag (6xHis) was ordered from Genscript Get A Quote

摘要

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 ... More

关键词

E. coli cell-based system, Hammer-head ribozyme, RNaseJ1, Reporter system, Ribozyme, glmS riboswitch