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Efficient in vitro transcription of plant nuclear tRNA Ser genes in a nuclear extract from tobacco cultured cells
Author(s) -
Yukawa Yasushi,
Sugita Mamoru,
Sugiura Masahiro
Publication year - 1997
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1046/j.1365-313x.1997.12040965.x
Subject(s) - transfer rna , primer extension , gene , transcription (linguistics) , biology , in vitro , microbiology and biotechnology , rna , arabidopsis , genetics , linguistics , philosophy , mutant
Summary 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.