Electroporated intact BY-2 tobacco culture cells as a model of transient expression study.
Transfer of foreign genes into plant cells can be accomplished by several methods: agrobacterium-mediated, microinjection, biolistic particle bombardment and electroporation. The last one is frequently used for transfection of plant protoplasts for transient gene expression. Electroporation is a simple procedure and allows transfecting a large number of cells at one time. Square wave-modulated porators are the most efficient for introducing expression cassettes into plant protoplasts. Based on a protocol developed by Wu & Feng (Plant Cell Reports, 1999, 18, 381-386), we optimized conditions for transfection of intact Nicotiana tabacum BY-2 cells using square wave-modulated electroporator. To simplify screening for transfected gene expression we used constructs with a GFP marker gene.
Kościańska
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E
Institute of Molecular Biology and Biotechnology, Adam Mickiewicz University, Poznań, Poland.
Wypijewski
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K
D002460:Cell Line; D002470:Cell Survival; D018274:Electroporation / Q000379:methods*; D018506:Gene Expression Regulation, Plant; D008856:Microscopy, Fluorescence; D014026:Nicotiana / Q000166:cytology / Q000235:genetics*; D014162:Transfection / Q000379:methods