title: Production of Homogeneous Recombinant RNA Using a tRNA Scaffold and Hammerhead Ribozymes.
abstract: Bacterial overproduction of recombinant RNA using a tRNA scaffold yields large amounts of chimeric RNA. For structural and functional characterizations of the RNA it is often necessary to remove the scaffold. Here we describe an efficient and facile method to release the RNA of interest from the tRNA scaffold by selective cleavage using cis-acting hammerhead ribozymes. After cleavage, the RNA of interest is purified to homogeneity using standard chromatographic and electrophoretic methods. Up to 5 mg of highly pure end-product RNA can be obtained from a single liter of bacterial culture.
authors.affiliation: Biophysical Chemistry, Institute for Molecules and Materials, Radboud University, Heyendaalseweg 135, 6525AJ, Nijmegen, The Netherlands.
authors.forename: Frank H T
authors.initials: FH
authors.lastname: Nelissen
authors.forename: Hans A
authors.initials: HA
authors.lastname: Heus
authors.forename: Sybren S
authors.initials: SS
authors.lastname: Wijmenga
mesh_terms: D005818:Genetic Engineering; D009690:Nucleic Acid Conformation; D012313:RNA / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D016337:RNA, Catalytic / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D012343:RNA, Transfer / Q000737:chemistry / Q000235:genetics*
chemical_list: D016337:RNA, Catalytic; C427204:hammerhead ribozyme; D012313:RNA; D012343:RNA, Transfer