title: Characterization of cells expressing RNA polymerase II tagged with green fluorescent protein: effect of ionizing irradiation on RNA synthesis.
abstract: OBJECTIVE
  To isolate and to characterize cells expressing RNA polymerase II tagged with green fluorescent protein for analyses of the effects of ionizing radiation on transcription in living cells.
  
  
  METHODS
  We introduced an alpha-amanitin-resistant mutation into a vector encoding the largest subunit of RNA polymerase II tagged with green fluorescent protein (GFP-pol). Cell lines stably expressing functional GFP-pol were isolated under selection with alpha-amanitin from a Chinese hamster cell line, CHO-K1, and a radiation-sensitive mutant CHO cell line, XR-1.
  
  
  RESULTS
  We tested the functionality of the fusion protein in vivo by determining RNA synthesis activity by incorporation of nucleoside analogues. Both CHO-K1 and XR-1 cells expressing GFP-pol had properties similar to those of their respective parental cell lines, indicating that GFP-pol is functional.
  
  
  CONCLUSIONS
  These stable lines might prove useful for analyses of the roles of transcription after ionizing radiation.
authors.affiliation: Radiation Effect Mechanisms Research Group, National Institute of Radiological Sciences, Inage-ku, Chiba, Japan.
authors.forename: Etsuko
authors.initials: E
authors.lastname: Hongo
authors.forename: Yoshie
authors.initials: Y
authors.lastname: Ishihara
authors.forename: Keiko
authors.initials: K
authors.lastname: Sugaya
authors.forename: Kimihiko
authors.initials: K
authors.lastname: Sugaya
mesh_terms: D053959:Alpha-Amanitin / Q000494:pharmacology; D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D004791:Enzyme Inhibitors / Q000494:pharmacology; D036681:Fluorescence Recovery After Photobleaching; D015971:Gene Expression Regulation, Enzymologic / Q000528:radiation effects*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D012313:RNA / Q000096:biosynthesis*; D012319:RNA Polymerase II / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000037:antagonists & inhibitors / Q000235:genetics*; D014158:Transcription, Genetic / Q000528:radiation effects*; D014965:X-Rays
chemical_list: D053959:Alpha-Amanitin; D004791:Enzyme Inhibitors; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D012313:RNA; D012319:RNA Polymerase II