title: Screening mutant libraries of T7 RNA polymerase for candidates with increased acceptance of 2'-modified nucleotides.
abstract: We present a screening assay based on fluorescence readout for the directed evolution of T7 RNA polymerase variants with acceptance of 2'-modified nucleotides. By using this screening we were able to identify a T7 RNA polymerase mutant with increased acceptance of 2'-methylseleno-2'-deoxyuridine 5'-triphosphate.
authors.affiliation: Department of Chemistry and Konstanz Research School Chemical Biology, University of Konstanz, Universitätsstrasse 10, 78457 Konstanz, Germany.
authors.forename: Vanessa
authors.initials: V
authors.lastname: Siegmund
authors.forename: Tobias
authors.initials: T
authors.lastname: Santner
authors.forename: Ronald
authors.initials: R
authors.lastname: Micura
authors.forename: Andreas
authors.initials: A
authors.lastname: Marx
mesh_terms: D019943:Amino Acid Substitution; D012321:DNA-Directed RNA Polymerases / Q000235:genetics / Q000378:metabolism*; D003856:Deoxyuracil Nucleotides / Q000737:chemistry / Q000378:metabolism; D015723:Gene Library; D057166:High-Throughput Screening Assays; D009930:Organic Chemicals / Q000737:chemistry; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D003856:Deoxyuracil Nucleotides; D009930:Organic Chemicals; C098798:SYBR Green II; D014764:Viral Proteins; C027078:deoxyuridine triphosphate; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases