ISSN:
1432-203X
Keywords:
Key words Plasmolysis
;
Electroporation
;
Gene transfer
;
β-Glucuronidase (GUS)
;
Green fluorescent protein (GFP)
;
Transient gene expression
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
Abstract This report describes the delivery of plasmid DNA containing either the β-glucuronidase (GUS) or the green fluorescent protein (GFP) reporter gene into intact plant cells of bamboo callus, lilium scales, and Nicotiana benthamiana suspension culture cells. By first plasmolyzing the tissues or cells with 0.4 m sucrose in the presence of plasmid DNA, electroporation effectively delivers plasmid DNA into the intact plant cells. Transient expression of the GUS gene, as revealed by histochemical assays, showed the presence of blue-staining areas in the electroporated tissues. A short exposure of cells to 2% DMSO (dimethyl sulfoxide) prior to plasmolysis elevated the level of transient GUS activity. When plasmid DNA containing a synthetic GFP gene was used, a strong green fluorescence was observed in N. benthamiana suspension culture cells that were subjected to plasmolysis and electroporation. These results suggest that plasmolysis brings the plasmid DNA into the void space that is in close vicinity to the plasmalemma, allowing electroporation to efficiently deliver the plasmid DNA into intact plant cells.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/s002990050590
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