title: Development of an efficient bi-directional promoter with tripartite enhancer employing three viral promoters.
abstract: We have developed a novel bi-directional promoter (FsFfCBD) by placing two heterogeneous core-promoters from the Figwort mosaic virus sub-genomic transcript promoter (FsCP, -69 to +31) and Cauliflower mosaic virus 35S promoter (CCP, -89 to +1) respectively on upstream (5') and downstream (3') ends of a tri-hybrid enhancer (FsEFfECE), in reverse orientation. The FsEFfECE domain encompasses three heterologous enhancer fragments from Figwort mosaic virus sub-genomic transcript promoter (FsE, 101 bp, -70 to -170), Figwort mosaic virus full-length transcript promoter (FfE, 196 bp, -249 to -54) and Cauliflower mosaic virus 35S promoter (CE, 254 bp, -343 to -90). The bi-directional nature of the FsFfCBD promoter (coupled to GFP and GUS) was established both in transient systems (onion epidermal cells and tobacco protoplasts) and transgenic plant (Nicotiana tabacum samsun NN) by monitoring the simultaneous expression of GFP and GUS employing fluorescence (for GFP) and biochemical (for GUS) based assays. In transgenic plants, the FsFfCBD promoter was found to be 6.8 and 2.5 times stronger than two parent promoters; Fs and FfC respectively. The bi-directional compound promoter FsFfCBD, composed of three heterologous enhancers with enhanced activity could become a valuable additional tool for efficient plant metabolic engineering and molecular pharming.
authors.affiliation: Division of Gene Function and Regulation, Institute of Life Sciences, Department of Biotechnology, Govt. of India, Nalco Square, Chandrasekherpur, Bhubaneswar 751 023, Odisha, India.
authors.forename: Sunita
authors.initials: S
authors.lastname: Patro
authors.forename: Indu B
authors.initials: IB
authors.lastname: Maiti
authors.forename: Nrisingha
authors.initials: N
authors.lastname: Dey
mesh_terms: D001709:Biotechnology / Q000379:methods; D017796:Caulimovirus / Q000235:genetics* / Q000378:metabolism; D004742:Enhancer Elements, Genetic; D005822:Genetic Vectors / Q000235:genetics* / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D019697:Onions / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011523:Protoplasts / Q000378:metabolism; D014026:Nicotiana / Q000235:genetics / Q000378:metabolism
chemical_list: D049452:Green Fluorescent Proteins