2S storage protein gene of Douglas-fir: characterization and activity of promoter in transgenic tobacco seeds.
To date a few sequences regulating expression of conifer seed-specific genes have been reported. To characterize Douglas-fir (Pseudotsuga menziesii [Mirb] Franco) 2S albumin storage protein genes, a genomic DNA sequence containing upstream promoter sequences was isolated by screening a Douglas-fir genomic library. Sequence analysis of the Douglas-fir gPm2S1 promoter revealed the presence of RY-repeated elements (GCATGC), and multiple E-box motifs (CANNTG) and ACGT-core elements, features characteristic of 2S storage protein genes in angiosperms. When fused to the GUS reporter gene, the 1.16 kb Douglas-fir 2S promoter sequence was sufficient to direct transient expression in both developing Douglas-fir embryos and maternally derived haploid megagametophytes. Analysis of this promoter construct in transgenic tobacco showed that expression was restricted to embryo and endosperm in developing seeds and was not detected in vegetative tissues of two-week-old seedlings. These results strongly suggest that both structural and regulatory elements as well as upstream signaling components controlling the expression of 2S albumin genes are highly conserved during evolution.
Chatthai
Malinee
M
Department of Biochemistry and Microbiology, Centre for Forest Biology, University of Victoria, Victoria, BC, Canada, V8W 3P6.
Forward
Benjamin S
BS
Yevtushenko
Dmytro
D
Stefanov
Ivan
I
Osuska
Lubica
L
Osusky
Milan
M
Misra
Santosh
S
D055516:2S Albumins, Plant; D000418:Albumins / Q000737:chemistry* / Q000235:genetics*; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D052179:Antigens, Plant; D001483:Base Sequence; D004247:DNA / Q000378:metabolism; D019143:Evolution, Molecular; D018014:Gene Transfer Techniques; D017343:Genes, Plant; D017930:Genes, Reporter; D005822:Genetic Vectors; D016678:Genome; D019684:Magnoliopsida / Q000378:metabolism; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000737:chemistry* / Q000235:genetics*; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic; D028224:Pseudotsuga / Q000378:metabolism*; D012639:Seeds / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid; D015398:Signal Transduction; D014026:Nicotiana / Q000235:genetics*; D014158:Transcription, Genetic; D019076:Transgenes
D055516:2S Albumins, Plant; D000418:Albumins; D052179:Antigens, Plant; D010940:Plant Proteins; D004247:DNA