Analysis of the 5' flanking region responsible for the endosperm-specific expression of a rice glutelin chimeric gene in transgenic tobacco

• Published in: Plant molecular biology Vol. 16; no. 1; pp. 49 - 58
• Main Authors: TAKAIWA, F, OONO, K, KATO, A
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer 1991
• Subjects: agrobacterium tumefaciens; arn; Base Sequence; Biological and medical sciences; Biotechnology; chimaeras; chimere; DNA, Recombinant; endosperm; endosperma; endosperme; Fundamental and applied biological sciences. Psychology; gene; Gene Expression Regulation; genes; Genetic engineering; Genetic technics; Glucuronidase - biosynthesis; Glucuronidase - genetics; gluteninas; glutenine; glutenins; Glutens - genetics; lycopersicon esculentum; Methods. Procedures. Technologies; Modification of gene expression level; Molecular Sequence Data; Nicotiana - genetics; Oryza - genetics; plantas transgenicas; plante transgenique; Plants, Toxic; Promoter Regions, Genetic; quimera; rna; Seeds - genetics; Transformation, Genetic; transgenic plants; Genetic mapping; Monocotyledones; Storage protein; RNA; Upstream sequence; Tissue specificity; Gene organization; Flanking sequence; Gene expression; Nicotiana tabacum; Oryza sativa; Gramineae; Dicotyledones; Angiospermae; Deletion; Regulatory sequence; Spermatophyta; Transgenic plant; Solanaceae; Glutelin; Hybrid gene
• ISSN: 0167-4412; 1573-5028
• DOI: 10.1007/bf00017916

The 5' upstream region of the rice storage protein type II glutelin gene was examined for its regulatory function in transgenic tobacco. Chimeric genes containing 5' flanking regions of the glutelin gene transcriptionally fused to the beta-glucuronidase (GUS) reporter gene were introduced into the tobacco genome by Agrobacterium tumefaciens-mediated gene transfer. The chimeric genes were expressed specifically in developing seeds, as opposed to leaves and stems, of the transgenic tobacco. Histochemical analysis revealed that the GUS activity was restricted to the endosperm tissue. A deletion series of the 5' flanking region was created from position -1329 to -74 relative to the transcriptional initiation site and similarly examined in transgenic tobacco. Measurement of GUS activity of the seeds from the transgenic plants bearing the chimeric genes indicated that the region between positions -441 and -237 was required for the temporal and endosperm-specific expression of the GUS activity in tobacco. RNA analysis by northern blotting confirmed the importance of the -441 to -237 region. Addition of up to 888 bp to the -441 deletion resulted in little increase in GUS activity, although all constructs expressing the GUS gene showed a similar tissue and temporal regulation pattern.