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.
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Acknowledgments
We thank Dr Hongquan Yang (Shanghai Jiaotong University, China) for the gift of plasmid pA7-GFP. We are grateful to Dr. Ryozo Sakiyama (University of Tokyo, Japan) for critical readings of this manuscript. This work was supported by the National Natural Science Foundation of China (30800082, 30571510, 30730078).
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Zhou, B., Zhao, X., Kawabata, S. et al. Transient expression of a foreign gene by direct incorporation of DNA into intact plant tissue through vacuum infiltration. Biotechnol Lett 31, 1811–1815 (2009). https://doi.org/10.1007/s10529-009-0080-8
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DOI: https://doi.org/10.1007/s10529-009-0080-8