title: Synthesis of guanosine 5'-conjugates and their use as initiator molecules for transcription priming.
abstract: We have synthesised two guanosine derivatives that are linked to biotinylated adenosine moieties by using two different strategies, one that includes synthetic steps on the solid phase and another one that is performed entirely in solution. The synthesised derivatives were shown to function as initiator molecules in transcription priming experiments. The incorporation efficiency was determined to be approximately 2%. Even though this value is rather low, the use of either molecule in selection experiments seems reasonable. Basically, RNA libraries with sequence complexities of 10(15) to 10(16) can be generated. Labelling of such a library with our initiator molecule would still produce 10(13) to 10(14) labelled/functionalised sequences, and thus sufficient sequence space for selection.
authors.affiliation: Ernst-Moritz-Arndt-Universität Greifswald, Institut für Biochemie, Bioorganische Chemie, Felix-Hausdorff-Strasse 4, 17487, Greifswald, Germany.
authors.forename: Jörn
authors.initials: J
authors.lastname: Wolf
authors.forename: Valeska
authors.initials: V
authors.lastname: Dombos
authors.forename: Bettina
authors.initials: B
authors.lastname: Appel
authors.forename: Sabine
authors.initials: S
authors.lastname: Müller
mesh_terms: D000241:Adenosine / Q000138:chemical synthesis / Q000737:chemistry; D001710:Biotin / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D017931:DNA Primers / Q000138:chemical synthesis / Q000737:chemistry*; D012321:DNA-Directed RNA Polymerases / Q000737:chemistry; D006151:Guanosine / Q000031:analogs & derivatives / Q000138:chemical synthesis* / Q000737:chemistry; D008968:Molecular Conformation; D012313:RNA / Q000737:chemistry; D014158:Transcription, Genetic
chemical_list: D017931:DNA Primers; D006151:Guanosine; D012313:RNA; D001710:Biotin; D012321:DNA-Directed RNA Polymerases; D000241:Adenosine