Efficient in vitro transcription of plant nuclear tRNA(Ser) genes in a nuclear extract from tobacco cultured cells.
A nuclear extract derived from tobacco cultured BY-2 cells supports RNA polymerase III-dependent transcription of Arabidopsis tRNA(Ser) genes. Primer extension analysis indicated that the transcription starts at 6 bp upstream from the 5' end of tRNA coding region. Procedures for nuclear extraction and in vitro reaction conditions have been optimized for tRNA transcription, which allows direct detection of de novo synthesized tRNA by gel electrophoresis. This improved in vitro system yields a mature-sized tRNA of 85 nucleotides from the Arabidopsis tRNA(Ser) gene, indicating that efficient processing of the pre-tRNA also occurs in the tobacco nuclear extract.
Yukawa
Y
Y
Center for Gene Research, Nagoya University, Japan.
Sugita
M
M
Sugiura
M
M
D002467:Cell Nucleus / Q000378:metabolism; D002478:Cells, Cultured; D002874:Chromosome Mapping; D003062:Codon; D010947:Plants, Toxic; D018749:RNA, Plant / Q000235:genetics*; D012362:RNA, Transfer, Ser / Q000235:genetics*; D014026:Nicotiana / Q000166:cytology / Q000235:genetics*; D014158:Transcription, Genetic
D003062:Codon; D018749:RNA, Plant; D012362:RNA, Transfer, Ser