Tissue specificity and developmental pattern of amorpha-4,11-diene synthase (ADS) proved by ADS promoter-driven GUS expression in the heterologous plant, Arabidopsis thaliana.
Amorpha-4,11-diene synthase (ADS) of Artemisia annua L. is a sesquiterpene cyclase that catalyzes the conversion of farnesyl diphosphate into amorpha-4,11-diene in the biosynthesis of the antimalarial artemisinin. To explore the mechanisms regulating the tissue-specific and developmental distributions of ADS, a full ADS promoter was generated using PCR, and fused to GUS for introduction into Arabidopsis thaliana. ADSpro::GUS fusion transcripts were organ-specific, mainly present in the anthers and trichomes of the green tissues of the juvenile leaves. This result was consistent with the ADS transcription pattern observed in A. annua as examined by RT-PCR. To determine the subcellular localization of ADS, an open reading frame (ORF) of ADS was fused to the green fluorescent protein (smGFP) gene and introduced into the A. thaliana protoplasts. GFP fluorescence was located exclusively in the cytosol, an indication that ADS is a cytosol-localized protein.
Kim
Soon-Hee
SH
Program in Applied Life Chemistry, Department of Agricultural Biotechnology, Seoul National University, Seoul, Korea.
Chang
Yung-Jin
YJ
Kim
Soo-Un
SU
D019883:Alkyl and Aryl Transferases / Q000235:genetics*; D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000201:enzymology* / Q000254:growth & development; D017931:DNA Primers; D035264:Flowers / Q000201:enzymology; D018507:Gene Expression Regulation, Developmental; D018506:Gene Expression Regulation, Plant; D008969:Molecular Sequence Data; D018515:Plant Leaves / Q000201:enzymology; D011401:Promoter Regions, Genetic; D020133:Reverse Transcriptase Polymerase Chain Reaction; D036226:Seedlings / Q000201:enzymology
D017931:DNA Primers; D019883:Alkyl and Aryl Transferases; C403714:amorpha-4,11-diene synthase