4.1 Article

Molecular cloning and characterization of GbDXS and GbGGPPS gene promoters from Ginkgo biloba

Journal

GENETICS AND MOLECULAR RESEARCH
Volume 12, Issue 1, Pages 293-301

Publisher

FUNPEC-EDITORA
DOI: 10.4238/2013.February.4.3

Keywords

Molecular cloning; GbDXS; GbGGPPS; Promoter; Cis-acting elements; Ginkgo biloba

Funding

  1. National Natural Science Foundation of China [31000904]
  2. Foundation for Innovative Research Groups of Natural Science of Hubei Province [2011CDA117]
  3. Natural Science Foundation of Hubei Province [2006ABA005, 2009CDB232]
  4. Technological Project of China Hubei Provincial Science & Technology Department [2004AA204B03]

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Ginkgolides are key pharmaceutical components in Ginkgo biloba leaves. 1-Deoxy-D-xylulose-5-phosphate synthase (GbDXS) and geranylgeranyl pyrophosphate (GbGGPPS) genes are critical genes involved in ginkgolide biosynthesis. In this study, the promoters of GbDXS and GGPPS, with 676 and 570 bp in length, respectively, were cloned by chromosome walking. The cis-elements of GbDXS and GbGGPPS promoters were predicted and analyzed by the plant cis-acting regulatory element (CARE) database. We found some major cis-elements in the sequence of GbDXS and GbGGPPS promoters. The GbDXS promoter has 3 TATA boxes, 10 CAAT boxes, 6 GATA boxes, and 1 I box. The GbGGPPS promoter has 1 TATA box, 6 CAAT boxes, 6 GATA boxes, and 4 I boxes. Furthermore, some stress-related cis-elements in the promoters of GbDXS and GbGGPPS were found to be light-regulated elements, including sequences over-represented in light-induced promoters (SORLIP1-AT), GATA box, and I box, a gibberellin-responsive element (WRKY), salicylic acid-induced (GT-1), cold-and dehydration-responsive (MYC-Core), and copper-inducible (CURE-Core). Further analyses of these cis-elements will aid in elucidating the molecular mechanisms regulating the expression of the GbDXS and GbGGPPS genes during ginkgolide accumulation in G. biloba.

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