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

BMC Molecular and Cell Biology

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