Transcriptional interference in transgenic plants.
When a promoterless marker gene is transformed into the plant genome using the Agrobacterium vector system, on average 30% of the T-DNA inserts produce gene fusions. This suggests that the T-DNA is preferentially integrated into transcribed regions. Here, we proposed that this transcriptional activity is responsible for some of the variation in expression frequently observed among independent transformants. Using hybrid gene constructions, we show that transcriptional readthrough into a downstream gene with opposite orientation substantially reduces expression of this gene both in transient expression and in transgenic plants. Furthermore, a poly(A) signal/terminator can block readthrough and restore the expression of the gene. Finally, enzymatic analysis of calli suggests that less variation in neomycin phosphotransferase II synthesis is observed when the gene is separated from plant DNA by promoter and terminator elements.
Ingelbrecht
I
I
Laboratorium voor Genetica, Universiteit Gent, Belgium.
Breyne
P
P
Vancompernolle
K
K
Jacobs
A
A
Van Montagu
M
M
Depicker
A
A
D016960:Agrobacterium tumefaciens / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics*; D015870:Gene Expression / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics*; D019868:Kanamycin Kinase; D010770:Phosphotransferases / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000235:genetics*; D010947:Plants, Toxic; D010957:Plasmids / Q000235:genetics; D011061:Poly A / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D012045:Regulatory Sequences, Nucleic Acid; D014026:Nicotiana / Q000235:genetics*; D014158:Transcription, Genetic / Q000235:genetics*
D004269:DNA, Bacterial; D011993:Recombinant Fusion Proteins; C036483:T-DNA; D011061:Poly A; D010770:Phosphotransferases; D019868:Kanamycin Kinase