title: Antisense RNA inhibition of beta-glucuronidase gene expression in transgenic tobacco plants.
abstract: Antisense RNA was used to specifically inhibit the expression of a GUS gene introduced in a transgenic plant. A tobacco transformant containing a single intact copy of the GUS gene and showing relatively high constitutive levels of GUS activity (GUS +) was re-transformed with an Agrobacterium Ti-derived binary vector containing an antisense version of this reporter gene. The sense and antisense GUS genes were each under the regulation of the CaMV 35S promoter. Re-transformed plants contained 1-5 copies of the antisense construct and all showed a greater than 90% reduction in GUS activity relative to the original GUS + plant. This reduction in GUS activity correlated closely with the levels of GUS enzyme and steady state GUS mRNA observed in these plants. The relatively low levels of sense and antisense GUS transcripts found in the re-transformed plants may indicate a rapid degradation of the RNA:RNA duplex in the cell.
authors.affiliation: Department of Biochemistry, University of Ottawa, Ont., Canada.
authors.forename: L S
authors.initials: LS
authors.lastname: Robert
authors.forename: P A
authors.initials: PA
authors.lastname: Donaldson
authors.forename: C
authors.initials: C
authors.lastname: Ladaique
authors.forename: I
authors.initials: I
authors.lastname: Altosaar
authors.forename: P G
authors.initials: PG
authors.lastname: Arnison
authors.forename: S F
authors.initials: SF
authors.lastname: Fabijanski
mesh_terms: D015870:Gene Expression; D005822:Genetic Vectors; D005966:Glucuronidase / Q000235:genetics* / Q000378:metabolism; D009692:Nucleic Acid Heteroduplexes / Q000378:metabolism; D010944:Plants / Q000235:genetics* / Q000378:metabolism; D010947:Plants, Toxic; D016372:RNA, Antisense / Q000235:genetics* / Q000378:metabolism; D014026:Nicotiana / Q000235:genetics / Q000378:metabolism; D014170:Transformation, Genetic
chemical_list: D009692:Nucleic Acid Heteroduplexes; D016372:RNA, Antisense; D005966:Glucuronidase