Expression of a self-processing, pathogen resistance-enhancing gene construct in Arabidopsis

Biotechnol Lett. 2005 Mar;27(6):435-42. doi: 10.1007/s10529-005-1884-9.

Abstract

A gene cassette, p35S-CNO, was designed to express three gene products driven by a single constitutive CaMV 35S promoter. The individual coding regions were linked in frame to produce a single polyprotein, using spacer sequences encoding a specific heptapeptide cleavage recognition site (ENLYFQS) for the nuclear-inclusion-a (NIa) proteinase of tobacco etch virus (TEV). The protein coding sequences used were: a Trichoderma harzinum endochitinase, a truncated NIa proteinase of TEV, and a wheat oxalate oxidase. When p35S-CNO construct was tested in Arabidopsis thaliana, the polyprotein was properly cleaved after translation and the products exhibited functional enzymatic activity in vivo.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabidopsis / genetics*
  • Blotting, Western
  • Chitinases / genetics
  • Chitinases / metabolism
  • Electrophoresis, Polyacrylamide Gel
  • Endopeptidases
  • Gene Expression Regulation, Plant
  • Genetic Vectors / genetics
  • Oxidoreductases / analysis
  • Oxidoreductases / genetics
  • Oxidoreductases / metabolism
  • Plant Diseases / genetics*
  • Plants, Genetically Modified / genetics*
  • Recombinant Proteins / genetics
  • Seedlings / enzymology
  • Subcellular Fractions
  • Triticum / genetics
  • Viral Proteins / genetics

Substances

  • Recombinant Proteins
  • Viral Proteins
  • Oxidoreductases
  • oxalate oxidase
  • Chitinases
  • Endopeptidases
  • nuclear inclusion protein a, mosaic viruses