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Identification of MazF Homologue in Legionella pneumophila Which Cleaves RNA at the AACU Sequence.

J Mol Microbiol Biotechnol. 2018;

Shaku M, Park JH, Inouye M, Yamaguchi Y,.

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Codon Optimization	… E coli BL21(DE3) and DH5α were used The mazF-lp and mazE-lp gene, whose codon usage was optimized for recombinant protein expression in E coli, were synthesized (Genscript) and cloned into pBAD24 [Guzman et al, 1995] and pET28a (Millipore), respectively …	Get A Quote

摘要

MazF is a sequence-specific endoribonuclease that is widely conserved in bacteria and archaea. Here, we found an MazF homologue (MazF-lp; LPO-p0114) in Legionella pneumophila. The mazF-lp gene overlaps 14 base pairs with the upstream gene mazE-lp (MazE-lp; LPO-p0115). The induction of mazF-lp caused cell growth arrest, while mazE-lp co-induction recovered cell growth in Escherichia coli. In vivo and in vitro primer extension experiments showed that MazF-lp is a sequence-specific endoribonuclease cleaving RNA at AACU. The endoribonuclease activity of purified MazF-lp was inhibited by purified MazE-lp. We found that MazE-lp and the MazEF-lp complex specifically bind to the palindromic sequence present in the 5'-u... More