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Updated: Aug 12, 2026

Transient Gene Expression in Tobacco using Gibson Assembly and the Gene Gun
Published on: April 18, 2014
Expression of a downstream gene from a bicistronic transcription unit in transgenic tobacco plants
S Iida1, O Mittelsten Scheid, M W Saul
1Department of Biological Science and Technology, Science University of Tokyo, Chiba, Japan.
Abstract:
We have constructed a set of plasmids carrying an artificial compact stop-start codon sequence, TGATGTAACATGA, between an upstream open reading frame, terminating at one of the stop codons, and a downstream kanamycin-resistance (KmR)-encoding gene (nptII) initiating at the second ATG. These plasmids were introduced into tobacco protoplasts by direct gene transfer. The efficiency of expression of the downstream nptII gene was measured by scoring the number of KmR transformants. With a closer distance between the functional stop and start codons, a tendency to less efficient expression of nptII was found. The integration and expression of both genes as a bicistronic transcription unit were verified by Southern- and Northern-blot analyses. A possible application of the compact stop-start codon sequence for insertional mutagenesis is discussed.
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