title: Syn5 RNA polymerase synthesizes precise run-off RNA products.
abstract: The enzyme predominantly used for in vitro run-off RNA synthesis is bacteriophage T7 RNA polymerase. T7 RNA polymerase synthesizes, in addition to run-off products of precise length, transcripts with an additional non-base-paired nucleotide at the 3'-terminus (N+1 product). This contaminating product is extremely difficult to remove. We recently characterized the single-subunit RNA polymerase from marine cyanophage Syn5 and identified its promoter sequence. This marine enzyme catalyses RNA synthesis over a wider range of temperature and salinity than does T7 RNA polymerase. Its processivity is >30,000 nt without significant intermediate products. The requirement for the initiating nucleotide at the promoter is less stringent for Syn5 RNA polymerase as compared to T7 RNA polymerase. A major difference is the precise run-off transcripts with homogeneous 3'-termini synthesized by Syn5 RNA polymerase. Therefore, the enzyme is advantageous for the production of RNAs that require precise 3'-termini, such as tRNAs and RNA fragments that are used for subsequent assembly.
authors.affiliation: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, 240 Longwood Avenue, Boston, Massachusetts 02115, USA.
authors.forename: Bin
authors.initials: B
authors.lastname: Zhu
authors.forename: Stanley
authors.initials: S
authors.lastname: Tabor
authors.forename: Charles C
authors.initials: CC
authors.lastname: Richardson
mesh_terms: D001435:Bacteriophages / Q000201:enzymology*; D000458:Cyanobacteria / Q000821:virology; D012321:DNA-Directed RNA Polymerases / Q000302:isolation & purification / Q000378:metabolism*; D004795:Enzyme Stability; D009711:Nucleotides / Q000378:metabolism; D012313:RNA / Q000096:biosynthesis*; D012343:RNA, Transfer / Q000096:biosynthesis; D014158:Transcription, Genetic; D014764:Viral Proteins / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D009711:Nucleotides; D014764:Viral Proteins; D012313:RNA; D012343:RNA, Transfer; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases