title: Comparative analysis of synthetic DNA promoters for high-level gene expression in plants.
abstract: We have designed two near- constitutive and stress-inducible promoters (CmYLCV9.11 and CmYLCV4); those are highly efficient in both dicot and monocot plants and have prospective to substitute the CaMV 35S promoter. We performed structural and functional studies of the full-length transcript promoter from Cestrum yellow leaf curling virus (CmYLCV) employing promoter/leader deletion and activating cis-sequence analysis. We designed a 465-bp long CmYLCV9.11 promoter fragment (-329 to +137 from transcription start site) that showed enhanced promoter activity and was highly responsive to both biotic and abiotic stresses. The CmYLCV9.11 promoter was about 28-fold stronger than the CaMV35S promoter in transient and stable transgenic assays using β-glucuronidase (GUS) reporter gene. The CmYLCV9.11 promoter also demonstrated stronger activity than the previously reported CmYLCV promoter fragments, CmpC (-341 to +5) and CmpS (-349 to +59) in transient systems like maize protoplasts and onion epidermal cells as well as transgenic systems. A good correlation between CmYLCV9.11 promoter-driven GUS-accumulation/enzymatic activities with corresponding uidA-mRNA level in transgenic tobacco plants was shown. Histochemical (X-Gluc) staining of transgenic seedlings, root and floral parts expressing the GUS under the control of CmYLCV9.11, CaMV35S, CmpC and CmpS promoters also support the above findings. The CmYLCV9.11 promoter is a constitutive promoter and the expression level in tissues of transgenic tobacco plants was in the following order: root > leaf > stem. The tobacco transcription factor TGA1a was found to bind strongly to the CmYLCV9.11 promoter region, as shown by Gel-shift assay and South-Western blot analysis. In addition, the CmYLCV9.11 promoter was regulated by a number of abiotic and biotic stresses as studied in transgenic Arabidopsis and tobacco plants. The newly derived CmYLCV9.11 promoter is an efficient tool for biotechnological applications.
authors.affiliation: KTRDC, College of Agriculture, University of Kentucky, Lexington, KY, 40546, USA, dipak.sahoo2@uky.edu.
authors.forename: Dipak Kumar
authors.initials: DK
authors.lastname: Sahoo
authors.forename: Shayan
authors.initials: S
authors.lastname: Sarkar
authors.forename: Sumita
authors.initials: S
authors.lastname: Raha
authors.forename: Indu B
authors.initials: IB
authors.lastname: Maiti
authors.forename: Nrisingha
authors.initials: N
authors.lastname: Dey
mesh_terms: D017360:Arabidopsis / Q000235:genetics* / Q000502:physiology; D017796:Caulimovirus / Q000502:physiology*; D035264:Flowers / Q000235:genetics / Q000502:physiology; D015870:Gene Expression; D018506:Gene Expression Regulation, Plant / Q000235:genetics*; D017930:Genes, Reporter; D019697:Onions / Q000235:genetics / Q000502:physiology; D010935:Plant Diseases / Q000276:immunology; D018517:Plant Roots / Q000235:genetics / Q000502:physiology; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011523:Protoplasts; D011994:Recombinant Proteins; D036226:Seedlings / Q000235:genetics / Q000502:physiology; D013312:Stress, Physiological; D014026:Nicotiana / Q000235:genetics / Q000502:physiology
chemical_list: D011994:Recombinant Proteins
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