Functional characterization of cis-elements conferring vascular vein expression of At4g34880 amidase family protein gene in Arabidopsis

PLoS One. 2013 Jul 2;8(7):e67562. doi: 10.1371/journal.pone.0067562. Print 2013.

Abstract

The expression of At4g34880 gene encoding amidase in Arabidopsis was characterized in this study. A promoter region of 1.5 kb on the upstream of the start codon of the gene (referred as AmidP) was fused with uidA (GUS) reporter gene, and transformed into Arabidopsis plant for determining its spatial expression. The results indicated that AmidP drived GUS expression in vascular system, predominately in leaves. Truncation analysis of AmidP demonstrated that VASCULAR VEIN ELEMENT (VVE) motif with a region of 176 bp sequence (-1500 to -1324) was necessary and sufficient to direct the vascular vein specific GUS expression in the transgenic plant. Tandem copy of VVE increased vascular system expression, and 5'- and 3'- deletions of VVE motif in combination with a truncated -65 CaMV 35S minimal promoter showed that 11bp cis-acting element, naming DOF2 domain, played an essential role for the vascular vein specific expression. Meanwhile, it was also observed that the other cis-acting elements among the VVE region are also associated with specificity or strength of GUS activities in vascular system.

MeSH terms

  • Agrobacterium tumefaciens / genetics
  • Amidohydrolases / genetics*
  • Amidohydrolases / metabolism
  • Arabidopsis / enzymology
  • Arabidopsis / genetics*
  • Arabidopsis Proteins / genetics*
  • Arabidopsis Proteins / metabolism
  • Codon, Initiator
  • Gene Expression Regulation, Plant*
  • Genes, Reporter
  • Glucuronidase / genetics
  • Glucuronidase / metabolism
  • Molecular Sequence Data
  • Plant Leaves / enzymology
  • Plant Leaves / genetics*
  • Plants, Genetically Modified
  • Promoter Regions, Genetic*
  • Protein Structure, Tertiary
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism

Substances

  • Arabidopsis Proteins
  • Codon, Initiator
  • Recombinant Fusion Proteins
  • Glucuronidase
  • Amidohydrolases
  • amidase

Grants and funding

The authors have no support or funding to report.