title: Determination of the half-life of chloroplast transcripts in tobacco leaves.
abstract: The amounts of specific transcripts that accumulate in chloroplasts are determined by the rates of synthesis and degradation of the transcripts. The 3' untranslated region of transcripts is a major determinant of the stability of transcripts in chloroplasts. The half-lives of specific transcripts can be determined by northern blot analysis of a time course of transcripts in detached tobacco leaves incubated with actinomycin D, a potent transcription inhibitor. This analysis may be applied to transcripts of endogenous genes or of transgenes introduced into the chloroplast genome in transplastomic plants. Sequence determinants of transcript stability can be identified by analysis of transplastomic plants containing constructs of the green fluorescent protein (gfp) reporter gene fused to the sequences of interest.
authors.affiliation: National Center for Genetic Engineering and Biotechnology, Klong Luang, Pathumthani, Thailand.
authors.forename: Sithichoke
authors.initials: S
authors.lastname: Tangphatsornruang
authors.forename: John C
authors.initials: JC
authors.lastname: Gray
mesh_terms: D020413:3' Untranslated Regions / Q000235:genetics; D001483:Base Sequence; D002736:Chloroplasts / Q000235:genetics*; D003609:Dactinomycin / Q000494:pharmacology; D017930:Genes, Reporter; D054628:Genome, Chloroplast / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics; D006207:Half-Life; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology; D045332:Photosystem II Protein Complex / Q000235:genetics; D018515:Plant Leaves / Q000235:genetics; D030821:Plants, Genetically Modified / Q000235:genetics; D020871:RNA Stability / Q000235:genetics*; D018747:RNA, Chloroplast / Q000235:genetics*; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012273:Ribulose-Bisphosphate Carboxylase / Q000235:genetics; D017423:Sequence Analysis, RNA; D014026:Nicotiana / Q000166:cytology / Q000235:genetics*
chemical_list: D020413:3' Untranslated Regions; D019384:Nucleic Acid Synthesis Inhibitors; D045332:Photosystem II Protein Complex; D018747:RNA, Chloroplast; D012336:RNA, Ribosomal, 16S; D049452:Green Fluorescent Proteins; D003609:Dactinomycin; C101625:RbcL protein, plastid; D012273:Ribulose-Bisphosphate Carboxylase