Transient expression of a foreign gene by direct incorporation of DNA into intact plant tissue through vacuum infiltration.
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.
Zhou
Bo
B
College of Life Science, Northeast Forestry University, 150040, Harbin, China.
Zhao
Xia
X
Kawabata
Saneyuki
S
Li
Yuhua
Y
D017360:Arabidopsis / Q000201:enzymology; D029741:Brassica rapa; D018744:DNA, Plant / Q000235:genetics*; D004274:DNA, Recombinant / Q000235:genetics; D015870:Gene Expression; D017343:Genes, Plant; D005821:Genetic Techniques; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000378:metabolism; D019697:Onions / Q000378:metabolism*; D019441:Plant Epidermis / Q000166:cytology; D013347:Subcellular Fractions / Q000378:metabolism; D014162:Transfection; D014618:Vacuum
D018744:DNA, Plant; D004274:DNA, Recombinant; D049452:Green Fluorescent Proteins