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Transfer RNA‐mediated suppression of amber stop codons in transgenic Arabidopsis thaliana
Author(s) -
Betzner Andreas S.,
Oakes Marie P.,
Huttner Eric
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.1997.11030587.x
Subject(s) - biology , arabidopsis thaliana , gene , reporter gene , transactivation , transgene , microbiology and biotechnology , genetics , sterility , arabidopsis , gene expression , stop codon , cytotoxic t cell , transfer rna , suppressor , rna , mutant , in vitro
Summary An artificial amber suppressor tRNA Leu gene ( sup L.) was physically linked to a mutated gus reporter gene, p35S‐gus (amL), which was inactivated by an amber stop codon (amL). Upon introduction into Arabidopsis thaliana , the presence of the sup L. gene was found to be correlated with cytotoxic effects observed during tissue culture and in mature plants. Those primary transformants that displayed cytotoxic symptoms were shown by X‐Gluc staining to express GUS as a result of amber stop codon suppression in vivo . Phenotypically normal lines were found by RT‐PCR to express sup L. GUS activity above background level was barely detectable in these plants, indicating a low level expression of sup L. However, the remaining suppressor activity was still sufficient to transactivate an amber‐mutated male sterility gene, pA9‐barnase (amL1) when combined within the same plant by crossing. The suppressor tRNA Leu gene may thus be used in transgenic plants for gene transactivation.