Transient expression of a foreign gene by direct incorporation of DNA into intact plant tissue through vacuum infiltration

Biotechnol Lett. 2009 Nov;31(11):1811-5. doi: 10.1007/s10529-009-0080-8. Epub 2009 Jul 18.

Abstract

We previously established a method to induce transient expression of foreign genes in intact plant tissue to detect the subcellular localization of proteins. Here, we have inserted a putative bZIP protein HY5 gene (SeqID: EU386772), isolated from the seedlings of turnips Brassica rapa L. subsp. rapa 'Tsuda,' and a receptor-like kinase gene AtRLK (SeqID: AY531551.1), isolated from Arabidopsis, into the plasmid pA7-GFP. We accomplished the direct incorporation of DNA into onion epidermal tissue by vacuum infiltration. By detecting GFP, which was fused with AtRLK or putative BrHY5, we determined that BrHY5 is located in the nucleus and AtRLK is located in the plasma membrane. This approach can be thus used to study the transient expression of foreign genes in intact tissue.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabidopsis / enzymology
  • Brassica rapa
  • DNA, Plant / genetics*
  • DNA, Recombinant / genetics
  • Gene Expression*
  • Genes, Plant*
  • Genetic Techniques*
  • Genetic Vectors / genetics
  • Green Fluorescent Proteins / metabolism
  • Onions / metabolism*
  • Plant Epidermis / cytology
  • Subcellular Fractions / metabolism
  • Transfection
  • Vacuum

Substances

  • DNA, Plant
  • DNA, Recombinant
  • Green Fluorescent Proteins