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Updated: May 24, 2026

RIBO-seq in Bacteria: a Sample Collection and Library Preparation Protocol for NGS Sequencing
Published on: August 7, 2021
The ordered processing of intervening sequences in 23S rRNA of Rhodobacter sphaeroides requires RNase J
1Institut für Mikrobiologie und Molekularbiologie, University of Giessen, Giessen, Germany.
Abstract:
The essential processing of ribosomal rRNA precursors requires concerted and sequential cleavages by different endo- and exoribonucleases. Despite long lasting investigations of these processes the exact order of steps remained elusive. Many bacteria perform additional rRNA processing steps by removing intervening sequences within the 23S rRNA. This leads to disintegration of the 23S rRNA and discontinuously assembled fragments within the ribosomes. The maturation of these fragments also requires successive cleavage events by different RNases. Our study reveals that the 5'-to-3' exoribonuclease RNase J is responsible for the final 5'-end maturation of all three 23S rRNA fragments in the α-proteobacterium Rhodobacter sphaeroides. Additionally the results show that 5'- and 3'-processing steps are closely coupled: mature 5'-ends are a strict prerequisite for the final 3'-trimming of the 23S rRNA fragments.
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